CN1900279A - Polynucleotide for inducing apoptosis and its coded polypeptide and use - Google Patents

Abstract

The present invention discloses a new kind of polynucleotide coding human protein with apoptosis inducing function, its coded polypeptide and the antibody to the polypeptide. The present invention also discloses the application of the new polynucleotides in extraneously expressing in host cell to induce apoptosis. The present invention also discloses the application of the polypeptide and antibody in preparing medicine for preventing and treating human body's apoptosis relevant diseases, especially in developing medicine for AIDS, neural retrograde affection and other autoimmunie diseases and tumors.

Description

The polynucleotide of cell death inducing and encoded polypeptides thereof and purposes

Technical field

The invention belongs to biological technical field, relate to the gene expression regulation field, specifically, the present invention relates to the proteic polynucleotide of people that the new coding of a class has the cell death inducing function, and encoded polypeptides, the antibody of polypeptide.The invention still further relates to the apoptotic application of external source induced expression in host cell of the new polynucleotide of this class.The invention still further relates to described polypeptide, antibody in the preparation prevention disease relevant with the human body cell apoptosis, especially developing as the purposes on the medicine of autoimmune disease such as acquired immune deficiency syndrome (AIDS), neural degeneration and tumour with treatment.

Background technology

The intravital cell of people is doomed to dead, some death is physiological, some death then is pathologic, the research of cells involved death process, become a focus of biology, medical research in recent years, up till now for this reason, people have known that the death of cell has dual mode at least, i.e. necrocytosis and apoptosis.Necrocytosis is a kind of cell death way that is realized already, apoptosis then is a kind of cell death way of being familiar with gradually in recent years, apoptosis is a kind of basic biological phenomena of cell, removes multicellular organism to play a part necessity in the unwanted or unusual cell.It plays an important role in the growth of stable and a plurality of systems of the evolution of organism, interior environment.Apoptosis is not only a kind of special necrocytosis type, and has important biological significance and complicated The Molecular Biology Mechanism.

Apoptosis is the process of the strict control of polygene.These genes are very conservative between kind, as Bcl-2 family, caspase family, oncogene such as C-myc, cancer suppressor gene P53 etc., along with the development of Protocols in Molecular Biology has had suitable understanding to the process of apoptosis of many kinds, but the apoptotic process precise mechanism still imperfectly understands up to now.

Why apoptosis becomes a focus of people's research, is decided by the substantial connection of apoptosis and clinical disease to a great extent.This relation not only shows apoptosis and Study on Mechanism thereof, has illustrated the pathogenesis of a big para-immunity disease, and can cause appearance, particularly apoptosis and extremely people's attention of the substantial connection between tumour and the acquired immune deficiency syndrome (AIDS) of disease new therapy thus.

1) the HIV virus infection causes CD4 ⁺It is to pass through Apoptosis Mechanism that the T cell reduces

HIV infects and causes that acquired immune deficiency syndrome (AIDS), its main pathogenesis are to destroy CD4 specifically after HIV infects ⁺The T cell makes CD4 ⁺T and relative immunodeficiency easily cause opportunistic infection and tumour, but how specificity is destroyed CD4 after the HIV infection ⁺The T cell? in recent years think CD4 ⁺The reason that T lymphocyte absolute number significantly reduces mainly causes by Apoptosis Mechanism.This has not only illustrated AIDS is CD4 ⁺The hypocellular major cause of T also is that the treatment research of AIDS has indicated an important probing direction simultaneously.

2) from the apoptosis angle, tumour be because due to apoptosis is obstructed

Tumour cell that it is generally acknowledged vicious transformation is because of growth out of control, and hyper-proliferative sees that from apoptotic angle the apoptosis mechanism of then thinking tumour is suppressed and can not normally carries out the result of necrocytosis removing.Therefore, designing the tumor treatment method from the apoptosis angle is exactly the apoptosis signal transduction system that rebuilds tumour cell, promptly suppresses the existence expression of gene of tumour cell, activates the expression of death gene.

3) apoptotic research will bring real breakthrough to autoimmune disease

Autoimmune disease comprises an intractable immunologic derangement of big class and the disease that causes, the bone-marrow-derived lymphocyte of autoreactivity T lymphocyte and generation antibody is the main immunopathogenesis mechanism that causes autoimmune disease, under the normal circumstances, the activation of immunocyte is a very complicated process.Under the hormesis of autoantigen, the immunocyte of identification autoantigen is activated, thereby obtains removing by apoptotic mechanism.But as this mechanism obstacle takes place, the removing of discerning the immunologically competent cell of autoantigen so will produce obstacle, causes the autoimmunization illness of lymphopoiesis.

4) neural degeneration

The acceleration of knowing the senile dementia nerve cell apoptosis at present produces.Alzheimer's disease (AD) is a kind of irreversible degeneration sacred disease, and the sudden change of amyloid precursor protein (APP), senilism albumen-1 (PS1) and senilism albumen-2 (PS2) causes familial alzheimer's disease (FAD).Studies have shown that PS has participated in the regulation and control of nerve cell apoptosis, expressing excessively of PS1, PS2 can strengthen the susceptibility of cell to apoptotic signal.Two member Bcl-xl of Bcl-2 gene family and Bcl-2 participate in the adjusting of pair cell apoptosis.

Since the generation of a lot of diseases is relevant with the apoptosis imbalance with development, it is logical so some disease being carried out the intervention of therapeutic apoptosis.Many genes are relevant with promotion or inhibition apoptosis of tumor cells, and therefore transgenosis inductor inner tumour cell apoptosis by all means is the new trial of research oncotherapy, and therapeutic value is immeasurable.On the contrary, just can treat some degeneration's diseases with strengthening cell to the tolerance of apoptosis, therefore, the discovery of apoptosis-related genes and mechanism research thereof are significant for the machine-processed gene therapy that reaches disease from now on of further discussion apoptosis.

Summary of the invention

The research of people's gene group is international focus at present, except that the method for large scale sequencing, also lacks the high flux screening that begins from functional study and has the method for the gene of certain function.Deficiency at this present situation and existing medicine or reagent the purpose of this invention is to provide the proteic polynucleotide NIAG1 of people, 2,3,4,5,6,7,8,9,10,11,12,13 that the new coding of a class has cell death inducing.

Another object of the present invention provides this class polynucleotide encoded polypeptide.

Another object of the present invention provides carrier and this class polynucleotide that contain these class polynucleotide and carrier transforms or the host cell of transduction.

Another object of the present invention provides the antibody of this class polynucleotide encoded polypeptide and the nucleic acid molecule that is used to detect.

Another object of the present invention provides the apoptotic application of external source induced expression in host cell of the new polynucleotide of this class.

Another object of the present invention provides produces these polynucleotide and the method for its encoded polypeptides and the purposes of this polynucleotide and encoded polypeptides thereof.

For achieving the above object, the present invention is by the following technical solutions:

In a first aspect of the present invention, novel isolating polynucleotide are provided, it comprises the proteic nucleotide sequence that coding has the cell death inducing function, and this nucleotide sequence is selected from: (a) contain SEQ ID NO:2 with coding, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, the polynucleotide of the polypeptide of the aminoacid sequence of SEQ ID NO:26 have the polynucleotide of at least 70% similarity; (b) coding contains the polynucleotide of polypeptide that aminoacid sequence with SEQ ID NO.2, SEQ ID NO.4, SEQ ID NO.6, SEQ ID NO.8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26 has the aminoacid sequence of at least 70% similarity; (c) with (a) or polynucleotide complementary polynucleotide (b).

Preferably, the polypeptide of this polynucleotide encoding has the aminoacid sequence of the group of being selected from down: SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26.

Preferably, the sequence of these polynucleotide is shown at least 85% similarity with the nucleotides sequence that is selected from down group: (a) coding region sequence or the full length sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25; (b) at least one sequence of the sequence of in genetic code degeneracy scope, mentioning in corresponding to (a); (c) with (a) or at least one sequence of the sequence complementary sequence hybridization of mentioning (b).

More preferably, the sequence of these polynucleotide is selected from coding region sequence or the full length sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25.

In a second aspect of the present invention, above-mentioned Nucleotide encoded polypeptide is provided, and it comprises the polypeptide with the aminoacid sequence in the group of being selected from down: SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26; Or the polypeptide that has similarity more than at least 90% with above arbitrary aminoacid sequence, or its conservative property variation polypeptide or its active fragments or its reactive derivative.

Preferably, this polypeptide is the polypeptide with aminoacid sequence of the group of being selected from down: SEQ ID NO:2, SEQ ID NO:4, SEQ IDNO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26.

In a third aspect of the present invention, the carrier that contains above-mentioned polynucleotide is provided, and, also provides the host cell that is transformed or transduce by above-mentioned polynucleotide by the host cell that this carrier transforms or transduces.

In a fourth aspect of the present invention, provide and aforementioned polypeptides specificity bonded antibody, the nucleic acid molecule that can be used for detecting also is provided, it contains 8-100 successive Nucleotide in above-mentioned arbitrary polynucleotide.

In a fifth aspect of the present invention, provide the apoptotic application of external source induced expression in host cell of above-mentioned polynucleotide.

In a sixth aspect of the present invention, provide above-mentioned polynucleotide and polypeptide to prevent and/or treat purposes in the medicine with human body cell apoptosis diseases associated in preparation.Preferably, prevent and/or treat in preparation relevant to the human body cell apoptosis the purposes in the medicine of autoimmune diseases such as acquired immune deficiency syndrome (AIDS), neural degeneration and tumour.

Aspect the present invention ground the 7th, a kind of pharmaceutical composition is provided, it contains the polypeptide and the pharmaceutically acceptable carrier of inducing the human body cell apoptosis function that have among the present invention of safe and effective amount.

In a eighth aspect of the present invention, the external detection method of a kind of autoimmune disease or tumour is provided, utilize above-mentioned antibody or nucleic acid fragment to detect the existence or the level of the polypeptide in host's sample.

Other aspects of the present invention since disclosing of the technology of this paper will be apparent to those skilled in the art.

As used herein, " isolating " is meant that material separates (if natural substance, primal environment promptly is a natural surroundings) from its primal environment.Do not have separation and purification as polynucleotide under the native state in the active somatic cell and polypeptide, if but same polynucleotide or polypeptide from native state, separate with common other materials that exist, then be separation and purification.Such polynucleotide may be the parts of a certain carrier, the part that also possible such polynucleotide or polypeptide are a certain composition, since carrier or composition are not the compositions of their natural surroundings, these polynucleotide or polypeptide remain isolating.

As used herein, " similarity " is meant and is used for describing the height that detects same DNA base between sequence and the target sequence or amino-acid residue order proportion in Nucleotide or the peptide sequence comparison process, it is a kind of direct quantitative relation, recently measure degree similar between nucleotide sequence or the peptide sequence by the same or analogous percentage of part, this similarity per-cent can calculate by the existing comparison method in this area, example has the comparison method FASTA program (Pearson between sequence in twos, W.R.and Lipman, D.J.1988.Improved tools for biological sequence comparison.Proc.Natl.Acad.Sci.85:2444-2448), blast program (Altschul, S.F., et al.1990 Basic local alignment search tool.J.Mol.Biol.215:403-410) etc., or Multiple Sequence Alignment Method CLUSTAL W (CORPET, F.1998 Multiple sequencealignment with hierarchical clustering.Nucleic Acids Res., 16:10881-10890) etc.Homologous sequence is meant the different sequences that form through divergent evolution from a certain common ancestor, can judge homology between aligned sequences according to similarity per-cent.When similarity degree is very high between gene or protein, represents that they have one section common evolution course, thereby judge that they can have similar biological function.When similarity degree, detect sequence and target sequence may be a homologous sequence than being easier to infer with at least 50%.Preferably, has at least 70% similarity degree; More preferably, has at least 85% similarity degree; Best, has at least 90% similarity degree.And when the similarity degree is lower than 20%, just be difficult to determine or can't determine at all whether it has homology.

Polynucleotide of the present invention comprise that its complementary strand can be dna form or rna form.Dna form comprises the DNA of cDNA, genomic dna or synthetic.DNA can be strand or double-stranded.DNA can be coding strand or noncoding strand.As used herein, " coding has the polynucleotide of the polypeptide of cell death inducing function " can be the polynucleotide that comprise this polypeptide of encoding, and also can be the polynucleotide that also comprise additional code sequence and/or non-coding sequence.With the NIAG1 encoded polypeptide is example, and the coding region sequence of encoding mature polypeptide can be identical with the coding region sequence shown in the SEQ ID NO:1 or the varient of genetic code degeneracy.As used herein, " genetic code degeneracy " is meant that an amino acid has the phenomenon of several codons.The varient of the genetic code degeneracy of NIAG1 encoded polypeptide refer to the encode Nucleotide of polypeptide in the present invention for example with SEQ ID NO:2, and the coding region sequence shown in this Nucleotide and the SEQ IDNO:1 has difference.Have the polypeptide of inducing the human body cell apoptosis function for other, can the rest may be inferred.

The invention still further relates to the varient of above-mentioned polynucleotide, its coding has the polypeptide of identical aminoacid sequence or fragment, analogue and the derivative of polypeptide with the present invention.The varient of these polynucleotide can be the allelic variant of natural generation or the varient that non-natural takes place.These nucleotide diversity bodies comprise and replace varient, deletion mutation body and insert varient.As known in the art, allelic variant is the replacement form of polynucleotide, and it may be replacement, disappearance or the insertion of one or more Nucleotide, but can be from not changing the function of its encoded polypeptides in fact.

The invention still further relates to and the complementary sequence hybridization of polynucleotide sequence of the present invention and two sequences between have at least 50%, preferably at least 70%, the polynucleotide of at least 80% homogeny more preferably.The present invention be more particularly directed under stringent condition the interfertile polynucleotide of the complementary sequence of polynucleotide sequence therewith.In the present invention, " stringent condition " is meant: (1) than hybridization under low ionic strength and the comparatively high temps and wash-out, as 0.2 * SSC, and 0.1%SDS, 60 ℃; Or (2) hybridization the time is added with denaturing agent, as 50% (v/v) methane amide, 0.1% calf serum/0.1%Ficoll, 42 ℃ etc.; Or (3) only at the homogeny between the two sequences at least more than 95%, be more preferably 97% and just hybridize when above.And the polypeptide of interfertile polynucleotide encoding has identical biological function and activity with polypeptide of the present invention.

Polynucleotide sequence of the present invention can obtain with this area existent method.These technology including, but not limited to: (1) is by hybridization technique DNA isolation sequence; (2) artificial chemical synthesising DNA sequence; (3) by the required polynucleotide of the extensive acquisition in construction cDNA library; (4) pcr amplification technology.

First method is to make up genomic library or cDNA library earlier, filters out goal gene or sequence by technology such as molecular hybridizations from genomic library or cDNA library then.When the biological gene group was smaller, this method is success easily; When the biological gene group is very big, make up difficulty of its complete genomic library, the quantities of removing to clone goal gene again from huge library is also very big.

Second method is by the long dna fragmentation of the once synthetic 100-200bp of the sequence that designs, and connects into complete gene with these synthetic fragment combination again.The price of the method for the gene order of this synthetic length is very expensive.This method be mainly used in synthetic as primer, connect the first grade nucleic acid fragment.

The third method is with usual method construction cDNA library, this area, repeatedly after the order-checking, in conjunction with bioinformatic analysis technology (Ota et al.Nat Genet.2004 Jan; 36 (1): 40-5), obtain purpose cDNA clone on a large scale.The bioinformatic analysis technology includes but not limited to BLAST or BLAT and the comparison of existing public database, as the refseq database etc.; With Phred algorithm assessment sequencing quality; ATGpr algorithm with the probability of occurrence that calculates transcription initiation codon ATG screens full length cDNA sequence etc.

The 4th kind of method method (Saiki, the et al.Science 1985 of round pcr DNA amplification/RNA; 230:1350-1354).The primer that is used for PCR can suitably be selected according to sequence information of the present invention disclosed herein, and available ordinary method is synthetic.Available ordinary method is as the DNA/RNA fragment by gel electrophoresis separation and purifying amplification.The method of advantageous applications of the present invention is that the amplification in mixing the cDNA library of two-step approach flux RT-PCR technology obtains a large amount of cDNA clones.Mix the cDNA library and comprise existing cDNA library and tumour library.

Gene of the present invention, the perhaps available ordinary method of mensuration of nucleotide sequence such as various dna fragmentations, as dideoxy chain termination (Sanger et al.PNAS, 1977,74:5463-5467); Also available commercial sequencing kit etc.In order to obtain the cDNA sequence of total length, order-checking need be carried out repeatedly.Sometimes need to measure a plurality of clones' cDNA sequence, just can be spliced into the cDNA sequence of total length.

Polypeptide of the present invention can be recombinant polypeptide, natural polypeptides, synthetic polypeptide, preferred recombinant polypeptide.Polypeptide of the present invention can be the product of natural purifying, or the product of chemosynthesis, or uses recombinant technology to produce from protokaryon or eucaryon host (as bacterium, yeast, higher plant, insect and mammalian cell).Polypeptide of the present invention can be glycosylated, also can be nonglycosylated.Polypeptide of the present invention can comprise or not comprise initial methionine residues.

The present invention also comprises fragment, derivative and the analogue of the people's protein polypeptide with the polynucleotide encoding of inducing the human body cell apoptosis function.Term " fragment ", " derivative " are meant basically with " analogue " and keep and the natural identical biological function or the active polypeptide of people's protein polypeptide of inducing the human body cell apoptosis function that have of the present invention.Polypeptide fragment of the present invention, derivative and analogue can be: one or more conservative or substituted polypeptide of non-conservation amino-acid residue (preferential conservative amino acid residue) (a) are arranged, and the amino-acid residue that replaces like this can be also can not encoded by genetic code, or (b) in one or more amino-acid residues, has a polypeptide of substituted radical, or (c) mature polypeptide and another compound (such as the compound that prolongs the polypeptide transformation period) merge formed polypeptide, or (d) additional aminoacid sequence is fused to this peptide sequence and the polypeptide that forms (as leader sequence or secretion sequence or be used for the sequence or the proteinogen sequence of this polypeptide of purifying).

Polypeptide of the present invention can be by conventional recombinant DNA technology, the protein polypeptide (Science, 1984 that utilize polynucleotide sequence of the present invention to express or produce having of reorganization to induce the human body cell apoptosis function; 224:1431).May further comprise the steps:

(1), or transforms or the transduction proper host cell with the expression vector that contains these polynucleotide with polynucleotide of the present invention (or its varient);

(2) host cell that culturing step (1) obtains in suitable medium;

(3) separation, the required protein polypeptide of purifying from substratum or cell.

Polynucleotide among the present invention and polypeptide preferably provide with isolating form, more preferably are purified to homogeneous.

The present invention also relates to comprise the carrier of polynucleotide of the present invention.Among the present invention, the polynucleotide sequence that coding has people's protein polypeptide of inducing the human body cell apoptosis function can be inserted in the recombinant expression vector.Term " recombinant expression vector " refers to bacterial plasmid well known in the art, phage, yeast plasmid, vegetable cell virus, mammalian cell virus, as adenovirus, retrovirus, and perhaps other carriers.The carrier of Shi Yonging can be a prokaryotic expression carrier in the present invention, it also can be carrier for expression of eukaryon, as the expression vector (Rosenberg that in bacterium, expresses based on T7, et al.Gene, 1987,56:125), the carrier for expression of eukaryon pcDNATM3.1/myc-hisB (-) of high expression level (Invitrogen) in mammalian cell, pcDNA3.1/V5-His-TOPO (Invitrogen below is abbreviated as pcDT).The preferred pcDT of the present invention, it can directly be connected with the PCR product and makes up carrier for expression of eukaryon, has improved the efficient of large-scale production greatly.As long as can duplicate in host and stablize, any plasmid and carrier can be used.A key character of expression vector is to contain replication orgin, promotor, marker gene and translation controlling elements usually.Making up the expression vector that contains polynucleotide sequence of the present invention and transcribe/translate control signal with method well-known to those having ordinary skill in the art gets final product.These methods comprise (Sambrook such as extracorporeal recombinant DNA technology, DNA synthetic technology, the interior recombinant DNA technology of body, et al.Molecular Cloning, a Laboratory Manual, Cold Spring Harbor Laboratory.New York, 1989).

Polynucleotide sequence of the present invention can be connected to effectively and instruct mRNA synthetic on the suitable promotor in the expression vector.The representative example of these promotors has: colibacillary lac or trp promotor; The PL promotor of lambda particles phage; Eukaryotic promoter comprises LTRs and some other known may command gene expression promoter in protokaryon or eukaryotic cell or its virus of CMV immediate early promoter, HSV thymidine kinase promoter, early stage and late period SV40 promotor, retrovirus.Expression vector also comprises ribosome bind site and the transcription terminator that translation initiation is used.Expression vector preferably comprises one or more selected markers, being provided for selecting the phenotypic character of transformed host cells, as being used for colibacillary tsiklomitsin or amicillin resistance or eukaryotic cell and cultivating green fluorescent protein (GFP), neomycin resistance and the Tetrahydrofolate dehydrogenase of usefulness.

The invention still further relates to the host cell that produces through genetically engineered with above-mentioned carrier or polynucleotide of the present invention.Carrier of the present invention and polynucleotide can be used to transform appropriate host cell, have the protein of inducing the human body cell apoptosis function so that it can be expressed.Host cell can be a prokaryotic cell prokaryocyte, as bacterial cell; Or eukaryotic cell such as low, as yeast cell; Or higher eucaryotic cells, as mammalian cell.Representative example has: intestinal bacteria; Vegetable cell; The insect cell of fruit bat S2 or Sf9; The zooblast of CHO, COS or Bowes melanoma cells; 293T, Hela cell etc.

When polynucleotide of the present invention are expressed, transcribe enhancing if will make when in carrier, inserting enhancer sequence in higher eucaryotic cells.Enhanser is the cis acting factor of DNA, and 10-300 base pair arranged usually, acts on promotor transcribing with enhancing gene.Example has: at the SV40 enhanser of 100-270 the base pair in replication origin downstream, at the polyoma enhanser in replication origin downstream and adenovirus enhanser etc.

Those of ordinary skill in the art knows how to select appropriate carriers, promotor, enhanser and host cell.

Can carry out with routine techniques well known to those skilled in the art with the recombinant DNA transformed host cell.When host cell was prokaryotic cell prokaryocyte such as intestinal bacteria, the competent cell that can absorb DNA can be collected at the exponential growth after date, uses CaCl ₂Method is handled, and used step is well-known in the art.Alternative is MgCl ₂Handle, also the method for available electroporation is handled.When the host is eukaryotic cell, can select following transfection method: coprecipitation of calcium phosphate method, conventional mechanical method such as microinjection, electroporation, liposome packing etc.The transformant that obtains can be cultivated with ordinary method, expresses polynucleotide encoded polypeptide of the present invention.Select suitable conventional substratum according to selected host cell, under the condition that is suitable for the host cell growth, cultivate.After host cell grows into suitable cell density, with appropriate means such as temperature inversion or chemical induction, induce the promotor of selection, cell is cultivated for some time again.

Recombinant polypeptide in the aforesaid method can wrap by in cell, extracellular or on cytolemma, express or be secreted into the extracellular.If desired, can utilize its physics, chemical separating and the purification of Recombinant polypeptide by various separation methods with other characteristics.These methods are well-known to those skilled in the art, handle as the renaturation of routine, handle the combination of (salt analysis method), centrifugal, the broken bacterium of infiltration, ultrasonication, super centrifugal, sieve chromatography (gel-filtration), adsorption chromatography, ion exchange chromatography, high performance liquid chromatography and other various liquid chromatography (LC) technology or these methods with protein precipitant.

The invention still further relates to any a part of homologous nucleic acid fragment with polynucleotide of the present invention.As used herein, the length of " nucleic acid fragment " contains 15 Nucleotide at least, is preferably at least 30 Nucleotide, is more preferably at least 50 Nucleotide, and best is at least 100 Nucleotide.This nucleic acid fragment is the dna sequence dna of chemosynthesis on the basis of nucleotide sequence information of the present invention normally.Above-mentioned nucleic acid fragment can be used for pcr amplification technology (as primer) and have the polynucleotide of inducing the human body cell apoptosis function to determine and/or to separate coding; Also can be used as the used probe of hybridization.Also can be used for the RNA perturbation technique.Part or all of polynucleotide of the present invention also can be used as probe stationary on microarray (Microarray) or DNA chip, is used for analyzing the differential expression and the gene diagnosis of tissue gene.The mark of probe can be used radio isotope, fluorescein or enzyme (as alkaline phosphatase) etc.

Polypeptide of the present invention can be directly as the pharmacological agent disease, as autoimmune diseases such as acquired immune deficiency syndrome (AIDS), neural degeneration and tumour etc.; Also can be used for screening proteic antibody, polypeptide or other part that promotes or resist the function with cell death inducing, for example, screening can be used for promoting or suppressing the antibody of proteic function of the present invention.Albumen of the present invention with the reorganization of expressing screens peptide library, is used to seek the peptide molecule that can promote or suppress proteic function of the present invention of therapeutic value.

Polypeptide of the present invention can use separately or use with suitable pharmaceutical carrier combination back.Composition comprises the polypeptide or the antagonist of safe and effective amount and does not influence the carrier and the excipient of effect of drugs.These carriers can be water, glucose, ethanol, salt, damping fluid, glycerine and their combination.Pharmaceutical composition can be with mode administration easily, as by in part, intravenously, intraperitoneal, intramuscular, subcutaneous, the nose or the route of administration of intracutaneous.The consumption that delivers medicine to the patient depends on many factors, as administering mode, person's to be treated healthiness condition and diagnostician's judgement.

Polypeptide of the present invention also can use by express these polypeptide at live body.For example patient's cell can carry out the genetically engineered operation by the gene at external use code book invention polypeptide, then engineering cell is offered the patient, makes engineering cell this peptide species of high expression level in vivo, thereby reaches the purpose of treatment.

The proteic polynucleotide of people with cell death inducing function also can be used for multiple therapeutic purpose.Can be used on and treat in the gene therapy technology owing to have the people's protein abnormal expression or the active disease that causes unusually of the function of cell death inducing.The people's albumen with cell death inducing function that the gene therapy vector (as virus vector) of reorganization can be designed to express variation suppresses the endogenic proteic activity of people with cell death inducing function.People's protein gene with cell death inducing function of reorganization also can be packaged in the liposome and be transferred in the cell.

Suppress the oligonucleotide (comprising sense-rna and DNA) of polypeptide mRNA of the present invention and nucleic acid also within the scope of the invention.Sense-rna and DNA and nucleic acid can be synthetic with this area existent method.In order to increase the stability of nucleic acid molecule, available several different methods is modified it, as increasing the sequence length of both sides, the connection between ribonucleoside phosphoric acid thioester bond or peptide bond.

Polypeptide of the present invention and fragment thereof, derivative, analogue or the cell of expressing them can be used as antigen and produce antibody.These antibody include but not limited to the antibody that monoclonal antibody, polyclonal antibody, chimeric antibody, single-chain antibody, Fab fragment and Fab expression library produce.The antibody of polypeptide of the present invention can be produced with preparation method for antibody well known in the art.Example has: monoclonal antibody can with hybridoma technology production (Kohler and Milstein.Nature, 1975,256:495-497).The available polypeptide immune animal of the present invention of the production of polyclonal antibody is as rabbit, mouse, rat etc.Multiple adjuvant can be used for the enhancing immunity reaction, includes but not limited to freund's adjuvant.The variable region bonded chimeric antibody in human constant region and inhuman source can be produced with existing technology (Morrison etal.PNAS, 1985,81:6851).The also available existing technology production of single-chain antibody (U.S.Pat No.4946778).

Antibody of the present invention can be used in the immunohistochemistry technology, detects the albumen with cell death inducing function in the living specimen.Can also be used for clinical diagnosis, treatment, therapeutic evaluation of the disease relevant etc. clinically with people's albumen with cell death inducing function.For example use labelled with radioisotope and polypeptide bonded monoclonal antibody of the present invention, inject then and follow the tracks of its position and distribution in the body, can be used as a kind of atraumatic diagnostic method and come the positioning tumor cell, or judge whether tumour cell shifts.Antibody among the present invention can also be used for the treatment of or prevent and the relevant disease of people's albumen with cell death inducing function.The antibody that gives suitable dosage can stimulate or block proteic generation of the people with cell death inducing function or activity.

The invention still further relates to quantitatively and detection and localization has the diagnostic testing process of the protein level of cell death inducing function.These tests are known in the art, and comprise that FISH measures and radioimmunoassay.The protein level that detects in the experiment with cell death inducing function, the disease that can have the importance of albumen in various diseases of cell death inducing function with laying down a definition and be used to diagnose albumen to work with cell death inducing function.

Proteic polynucleotide with cell death inducing function can be used for having the diagnosis and the treatment of the protein related diseases of cell death inducing function.Aspect diagnosis, the proteic polynucleotide with cell death inducing function can be used for detecting have the cell death inducing function proteic expression whether, or under morbid state, have the abnormal exprssion of cell death inducing function.As the proteic dna sequence dna with cell death inducing function can be used for the hybridization of biopsy specimen is had with judgement the proteic abnormal expression of cell death inducing function.Hybridization technique is the disclosed mature technology in this area, comprises Southern blotting, Northern blotting, in situ hybridization etc., and relevant test kit can obtain from commercial channels.Part or all of polynucleotide of the present invention can be used as probe stationary on microarray (Microarray) or DNA chip, is used for analyzing the differential expression and the gene diagnosis of tissue gene.Carry out RNA-polymerase chain reaction (RT-PCR) amplification in vitro with the special primer of the albumen with cell death inducing function and also can detect proteic transcription product with cell death inducing function.

The sudden change that detection has the protein gene of cell death inducing function also can be used for diagnosing the relevant disease of albumen with cell death inducing function.Mutant form with protein gene of cell death inducing function comprises that the point mutation compared with the proteic dna sequence dna with cell death inducing function of normal wild type, transposition, disappearance, reorganization and other are any unusual etc.Existing technology in available this area such as Southern blotting, dna sequence analysis, PCR and in situ hybridization detect sudden change.In addition, sudden change might influence proteic expression, therefore can judge indirectly that with Northern blotting, Westen blotting gene has or not sudden change.

In gene NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 healthy tissues that all are used in experiment, fetal tissue, the tumor tissues expression is arranged all, illustrate that it is the important apoptosis-related genes of human body self; Expression amount just has difference in different tissues, and the degree difference of its performance function in different tissues is described.

Below embodiment of the present invention are further described.

The present invention carries out the retrieval of people's Unknown Function predicted gene by the refseq database to NCBI, obtain people's unknown function gene order, further utilizing the Human_est database to carry out sequence by the BLASTn method proofreaies and correct, according to the sequences Design gene specific primer that obtains after proofreading and correct, from mix people's tissue cDNA library, obtain the coding region cDNA fragment of goal gene by the amplification of two-step approach flux RT-PCR technology.This coding region cDNA fragment and pcDT recombination to construct carrier for expression of eukaryon.Adopt the gene induced apoptotic function among the renilla luciferase reporter gene method detection the present invention of pRL family.The pRL vehicle group is expressed renilla luciferase with becoming second nature, and under the identical situation of other transfection conditions, the active power of renilla luciferase can reflect the influence of gene to be checked for cell state, as cell number, cell survival rate etc.To compare the active gene that obviously reduces of renilla luciferase with the pcDT empty plasmid picks out.Behind the independent transfectional cell of these genes, utilize LIVE/DEAD  Viability/Cytotoxicity Kit (L-3224) dyestuff pair cell to dye, further observe and verify the cell survival state.Evidence in this test kit, Calcein-AM is a kind of endochylema fluorescent marker, no fluorescence own, the water-soluble green fluorescence material that the catalysis of cell lactonase generates behind the infiltration cell is difficult for appearing cell, is used for viable cell dyeing.EthD-1 can not enter viable cell but can enter the damaged dead cell of film, and is incorporated on the DNA, produces a kind of red fluorescence, is used for dead cell stain.The survival condition that can reflect cell by the ratio of the red green fluorescence of fluorescence microscope.And then detect the influence of selected gene pairs cell by fluidic cell experiment, and all obtain positive findings, what all degree was different has apoptosis-induced effect to institute's transfectional cell.Experiment shows that polypeptide of the present invention has remarkable, stable cell death inducing effect.

Owing to adopted above technical scheme, the present invention has following advantage:

1, provide mass-producing to clone and screen the technology platform of new gene.

2, human new functional gene NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 cDNA sequence and coded polypeptide thereof are provided;

3, find that first human new functional gene NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 has the effect of cell death inducing, and efficient, stable;

4, NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 expresses at the most normal cells of body, illustrates that it is the relevant regulatory molecule of self important apoptosis.

5, based on 4 above-mentioned advantages, the present invention is the apoptosis mechanism of further studying, and exploitation treatment is established necessary base as the novel drugs of autoimmune disease such as acquired immune deficiency syndrome (AIDS), neural degeneration and tumour etc. for starting new clinical diagnosis, therapeutic evaluation and prognostic indicator.

Description of drawings

The structure synoptic diagram of Fig. 1, carrier for expression of eukaryon pcDT-NIAGx

(NIAGx is selected from NIAG1,2,3,4,5,6,7,8,9,10,11,12,13)

Fig. 2, pRL family renilla luciferase gene reporter plasmid structure iron

Fig. 3, NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 heterogenous expressions are to the influence of pRL luciferase expression

The influence of Fig. 4, NIAG1,2,3,4,5,6,7,8,9,10,11,12, the survival of 13 heterogenous expression pair cells (light field: 10 *)

The influence of Fig. 5, NIAG1,2,3,4,5,6,7,8,9,10,11,12, the survival of 13 heterogenous expression pair cells (Calcein AM and EthD-1 dyeing: 10 *)

Fig. 6, NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 heterogenous expression cell death inducings (flow cytometry experiment)

Embodiment

Below in conjunction with specific embodiment, further set forth the present invention.These embodiment only are used to the present invention is described and are not used in and limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, usually according to normal condition, condition described in " molecular cloning experiment guide " (chopsticks such as the third edition [U.S.] Sa nurse Brooker in 2002, Science Press), or the condition of advising according to manufacturer.

Embodiment 1, two-step approach flux RT-PCR technology amplifying target genes

(1) the refseq database to NCBI carries out the retrieval of people's Unknown Function predicted gene, obtain people's unknown function gene order, and utilize the Human_est database to carry out sequence by the BLASTn method and proofread and correct, the sequence that finally obtains is set at down the group sequence: SEQID NO.1, SEQ ID NO.3, SEQ ID NO.5, SEQ ID NO.7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ IDNO:25.According to this type of sequences Design gene NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 special primer:

The gene title	Upstream primer (5 '-3 ')	Downstream primer (5 '-3 ')
			NIAG1 NIAG2 NIAG3 NIAG4 NIAG5 NIAG6 NIAG7 NIAG8 NIAG9 NIAG10 NIAG11 NIAG12 NIAG13	gtcgacgctgcttcgttg tgtcgtgaggctgaggtgcag cctcggttaaacactggtcgttc tgtgtccgtcgccatgacag acttcatcgtcatgcagagcgtc ggaggtcgctgctatgatccg cctcggttaaacactggtcgttc tgtcccatgaggctgccc cattgatgcacccattccagtg atgccaggtcgttggaggt gggtggccgaagcagaagac gcaaccatcaatcccgtctcc ctgagggaacgctaagtagtgtgtc	gcctcctggagtgtggtggtc gtgaaacgtggttagtctcgtaaaatg ttctgtcactcttactaggattcgcc tggttgagtggcaggtgagg ccgtcactccgtgtgcgc gccttactggatcacacacagtttc ttctgtcactcttactaggattcgcc tccctaatagtagcgattcaggctc tctccaggtctcggtgtagcg gaggagggagctacaactcg ggcagactctcaagatattcacaacc ttgggaccaagtagtgccatttac gaagtctacatggtgttaattgctgg

(2) use above-mentioned primer, in existing cDNA library and tumour library, select template, carry out just expanding by the express spectra of goal gene.Existing library comprises 12 kinds of human normal tissues (heart, pancreas, testis, ovary, prostate gland, colon, small intestine, skeletal muscle, thymus gland, lymphoglandula, tonsilla, white corpuscle); 6 kinds of people's tumor tissues (lung cancer, carcinoma of the pancreas, ovarian cancer, prostate cancer, colorectal carcinoma, mammary cancer); With the cDNA library of 8 kinds of fetuses group long-pending (tire lung, fetal rhythm, tire liver, tire spleen, tire kidney, tire brain, tire skeletal muscle, tire thymus gland) (Clonetch, K1420-1,1241-1).It is as follows just to expand reaction conditions:

50 μ l PCR reaction:

CDNA mixing storehouse 5 ', 3 ' primer, 10 * Pyrobest buffer 2.5mM dNTPs Pyrobest distilled water

Each 1.0 μ l final concentration is that 2pmol/ μ l 5 μ l 4 μ l 1 μ l complements to 50 μ l

PCR extends the long segment of time according to the expansion goal gene, increases by the principle of 50sec/Kb:

94 ℃ 5 minutes; 94 ℃ 30 seconds, 64 ℃ 30 seconds, 72 ℃ of 50sec/Kb; 72 ℃ 10 minutes4 ℃ of ∞

28 circulations

The thing of just expanding production is purified to 30 μ l, with primers a large amount of in the removal PCR reaction system and dNTPs etc., and concentrates whole system, obtains the secondary amplification bank of corresponding target gene sequences, as two templates that expand (the big expansion).

(3) with the purified product in (2) as template, respectively each goal gene is carried out two expansions, reaction conditions is as follows: 50 μ l PCR (each gene) reaction:

One expands purified product 5 ', 3 ' primer 10 * Ex-Taq buffer 2.5mM dNTP Ex-Taq distilled water

1.6 being 2pmol/ μ l 5 μ l 4 μ l 0.5 μ l, μ l final concentration complements to 50 μ l

PCR extends the long segment of time according to the expansion goal gene, increases by the principle of 50sec/Kb:

94 ℃ 5 minutes; 94 ℃ 30 seconds, 64 ℃ 30 seconds, 72 ℃ of 50sec/Kb; 72 ℃ 10 minutes4 ℃ of ∞

30 circulations

The PCR product that obtains is got sample electrophoresis on the 10 μ l, selects the PCR product of amplified band, carries out equal-volume purifying (40 μ l).The gene that amplifies non-single band by the two-step pcr reaction reclaims test kit with Qiagen glue and cuts glue recovery purpose fragment.The result of amplification shows, all there are gene NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 cDNA in the cell of these tissues, illustrate that NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 has produced gene NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 transcription product in the cell of these tissues, wider expression map is arranged, in multiple tissue, participate in the adjusting of transcription factor.

Embodiment 2, goal gene Construction of eukaryotic

With two expansion purified product and carrier for expression of eukaryon pcDNA3.1/V5-His-TOPO (Invitrogen is abbreviated as pcDT), carry out ligation according to the condition of test kit manufacturer suggestion.Connect product electric shocking method transformed into escherichia coli DH5 α, conversion product is grown containing on the solid LB plate culture medium of penbritin, select the monospecific polyclonal bacterium colony of growth, extract plasmid, cut with the EcoRI enzyme, enzyme is cut product and is identified with agarose gel electrophoresis, has selected and has inserted segmental positive colony, select correct forward by order-checking (ABI PRISM 3700 DNA analysis instrument) and insert clone, called after NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 separately.

Collect nutrient solution simultaneously, analyze protein precipitation, obtain NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 polypeptide with SDS-PAGE.

NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 analysis of protein results show: NIAG1,2,3,4,5,6,7,8,9,10,11,12, following group of sequence of 13 protein sequences: SEQ ID NO.2, SEQ ID NO.4, SEQ ID NO.6, SEQ ID NO.8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, shown in the SEQ ID NO:26.

Embodiment 3, renilla luciferase reporter gene method are measured the effect of goal gene cell death inducing

With goal gene and the renilla luciferase reporter gene carrier cotransfection human embryo kidney 293T of pRL family cell, measure the influence of goal gene by the activity that detects renilla luciferase, as cell number, cell survival rate etc. for cell state.To compare the active genescreen that obviously reduces of renilla luciferase with the pcDT empty plasmid comes out.The concrete operations step is as follows:

(1) cell cultures: with 1.2 * 10 ⁴Individual 293T cell (ATCC Number:CRL-11268) is with DMEM (Dulbecco ' the s modified Eagle ' s medium) substratum (Hyclone that contains 10% foetal calf serum, SH0022.02) be layered on 96 porocyte culture plate (Costar, 3599) go up (100 μ l nutrient solution/hole), at 5%CO ₂, 37 ℃ cell culture incubator (SANYO, MCO-15AC) the middle cultivation 24 hours.

(2) preparation transfection working fluid: with 2.5 μ l physiological saline dilution 20ng pRL family's renilla luciferase reporter gene carrier and 80ng gene to be checked, mixing gently, room temperature placement; Dilute 0.04 μ l VigoFect transfection reagent (prestige lattice Lars biotechnology (Beijing) company limited) with 2.5 μ l physiological saline equally, mixing gently, room temperature was placed 5 minutes; The VigoFect transfection reagent of dilution is dropwise added in the gene to be checked and reporter gene mixed solution of dilution, and mixing is to be the transfection working fluid gently, and room temperature was placed 15 minutes.

(3) transfection: with transfection working fluid mixing gently, dropwise be added in the 96 porocyte culture plates of completing cell in (1), place 37 ℃, 5%CO ₂Cell culture incubator in cultivated 24 hours.

(4) detect: transfection is after 24 hours, and (Eppendorf Centrifuge 5810R) centrifugal 5 minutes, abandons supernatant with 800 rev/mins with the cell of 96 orifice plates.Every hole adds 40 μ l cell pyrolysis liquids, places-80 ℃ of refrigerators more than 1 hour 96 orifice plates behind the mixing.Return to room temperature after 96 orifice plates are thawed from-80 ℃, every hole is drawn 10 μ l cell pyrolysis liquids and is moved to white enzyme plate, and (Genios Pro Tecan) detects the renilla luciferase activity with the microwell plate microplate reader.Renilla luciferase substrate consumption is every hole 25 μ l.

The pRL carrier is with renilla luciferase gene and basic expression promoter coupling, and constructive expression's renilla luciferase is provided.The change of condition can influence the expression of renilla luciferase in the experimentation, and these factors comprise cell survival situation (cell number, cell health situation etc.), transfection efficiency, lysis efficient and detection efficiency etc.Under the prerequisite of experimental system error, the transfection of same 96 orifice plate and detection efficiency are approximate substantially, therefore, the active obvious reduction of renilla luciferase can reflect the change of cell survival situation, we with the pcDT plasmid in contrast, will with the pRL plasmid co-transfection after cause that the active genescreen that obviously reduces of renilla luciferase comes out.

The result is as shown in Figure 3: gene NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 and pcDT relatively, obviously reduced the renilla luciferase activity.Show that gene NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 can change the survival condition of transfectional cell.

Embodiment 4, utilize dyestuff observation of cell survival condition

Behind the independent transfection 293T of the gene that filters out among the embodiment 3 cell, utilize LIVE/DEAD  Viability/Cytotoxicity Kit (Molecular Probe, L-3224) pair cell dyes, operate by product description is described, observation of cell survival condition (comprising light field and fluorescence excitation state) under inverted fluorescence microscope (Zeiss, Axiovert 200M).The concrete operations step is as follows:

This experiment is with 96 porocyte culture plates (Costar, 3599) operations, and each gene to be checked is provided with 3 parallel repeating holes, respectively with pcDT empty plasmid and Bax plasmid as empty carrier and positive control.Consumption in the following step is the single hole consumption.

(1) cell cultures: with 1.2 * 10 ⁴Individual 293T cell is layered on (100 μ l nutrient solution/hole) on the 96 porocyte culture plates with the DMEM substratum that contains 10% foetal calf serum, puts 37 ℃, 5%CO ₂Cell culture incubator in cultivated 24 hours.

(2) preparation transfection working fluid: with 2.5 μ l physiological saline dilution 160ng gene to be checked, mixing gently, room temperature placement; Dilute 0.04 μ l VigoFect transfection reagent with 2.5 μ l physiological saline equally, mixing gently, room temperature was placed 5 minutes; The VigoFect transfection reagent of dilution dropwise is added in the cdna solution to be checked of dilution, and mixing is to be the transfection working fluid gently, and room temperature was placed 15 minutes.

(3) transfection: with transfection working fluid mixing gently, dropwise be added in the 96 porocyte culture plates of completing cell in (1), mixing is put 37 ℃, 5%CO gently ₂Cultivated 24-48 hour in the cell culture incubator.

(4) microscopy: after transfection 24-48 hour, change with Zeiss microscope observing cell form.Plasmid to be checked compares with pcDT and Bax respectively.The result as shown in Figure 4, transfection NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 genes are after 30 hours, institute's cells transfected base state is relatively poor, volume-diminished connect to disappear, caryoplasm concentrates look dark, and a side relatively.

(5) dyeing: diluting 10 μ l 2mM EthD-1 (dead cell easily dyes) storage liquid to concentration with 5ml PBS is 4 μ m, fully mixing; With 5ml PBS solution dilution 2.5 μ l 4mM Calcein AM (viable cell dyestuff) storage liquid, abundant mixing; Two kinds of dyestuff final concentrations are respectively 1 μ M Calcein AM and 2 μ M EthD-1.Discard the nutrient solution of transfectional cell, use the PBS (37 ℃) of incubation to wash twice.Add 50 μ l and dilute good dyestuff, put 37 ℃, CO ₂Hatched in the incubator 30 minutes to 1 hour.

(6) microscopy: discard dyestuff,, add 100 μ l PBS, observe with the Zeiss inverted fluorescence microscope with PBS washing one time.Calcein AM excitation wavelength is 485 ± 10nm, and emission light is 530 ± 12.5nm; The EthD-1 excitation wavelength is 530 ± 12.5nm, and emission light is 645 ± 20nm.

The result as shown in Figure 5, gene NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 and empty carrier relatively, the existing state of institute's transfectional cell is obviously not good in the same field of view, erythrophil (EthD-1 staining cell, rubescent look fluorescence) more, and green transfect cell (Calcein AM staining cell, green-emitting fluorescence) less, approaching with positive control Bax effect, illustrate that NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 can obviously change the cell survival state, and inducing institute's transfectional cell necrosis or apoptosis in varying degrees.

Embodiment 5, fluidic cell experiment

Behind the independent transfection 293T of the gene that filters out among the embodiment 3 cell, the inducing cell that utilizes flow cytometer to detect plasmid to be checked is transferred the function of dying.The concrete operations step is as follows:

(1) cell cultures: with 293T cell (1 * 10 ⁵) be layered on the 24 porocyte culture plates (Costar, 3599) with the DMEM substratum that contains 10% foetal calf serum, put 37 ℃, 5%CO ₂Cell culture incubator in cultivated 16 hours.

(2) preparation transfection working fluid: with 8 μ l physiological saline dilution 500ng plasmid to be checked, mixing gently, room temperature is placed: dilute 0.16 μ l VigoFect transfection reagent with 8 μ l physiological saline equally, mixing gently, room temperature placement 5 minutes; The VigoFect transfection reagent of dilution dropwise is added in the plasmid solution to be checked of dilution, mixing gently, room temperature was placed 15 minutes.

(3) transfection: with transfection working fluid mixing gently, dropwise be added in the Tissue Culture Plate (500 μ l nutrient solution/hole), mixing is put 37 ℃, 5%CO gently ₂Cultivated 24-48 hour in the incubator.

(4) preparation single cell suspension, flow cytometer detects

24 and 48 hours harvested cells after the transfection.The sucking-off substratum, add 100 μ l pancreatin (Hyclone, SH30042.01), 37 ℃ of digestion 3 minutes adds 200 μ l again and contains in the DMEM substratum of 10% foetal calf serum and pancreatin, pressure-vaccum makes formation unicellular for several times, move into streaming pipe (FALCON, 352052), 1500 rpms of (Eppendorf Centrifuge, 5810R) room temperature is centrifugal 5 minutes, supernatant discarded.Add 3ml PBS re-suspended cell, centrifugal 5 minutes of 1500 rpms of room temperatures, supernatant discarded.Repeat above-mentionedly to wash cell once, and be resuspended in 200 μ l Binding buffer (10mM Hepes, 140mM NaCl, 1mM MgCl ₂, 5mM KCl, 2.5mM CaCl ₂), add final concentration and be 1 μ g/ml FITC-Annexin-V (Beijing Bao Sai Bioisystech Co., Ltd, CX1001-2), room temperature lucifuge reaction 30 minutes, add final concentration again and be 1 μ g/ml PI (Beijing Bao Sai Bioisystech Co., Ltd, CX1001-2).Detection use flow cytometer (FACSCalibur, BD, USA).The Annexin-V green-emitting fluorescence of FITC mark can specificity be incorporated into the distinctive phosphatidylserine that turns up outside cytolemma of apoptotic cell, and PI can penetrate the cytolemma of dead cell, and then nucleus is dyed redness.The cell of Annexin-V and PI jack to jack adapter is viable cell, and all the other are dead cell.

The result as shown in Figure 6, no matter be transfection 24 hours or 48 hours, NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 and empty carrier positive findings is more all arranged, similar to positive control Bax effect, be each gene action degree varies, and the effect of 48 hours cell death inducings is more more remarkable than 24 hours.Illustrate NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 in varying degrees inducing cell transfer and to die.

Embodiment 6, Antibody Preparation

Antigen is selected NIAG1,2,3,4,5,6,7,8,9,10,11,12,13 albumen total lengths or the partial peptide section of prokaryotic cell prokaryocyte or eukaryotic cell expression for use, also can synthesize polypeptide as antigen.

Polyclonal Antibody Preparation: immune animal is selected bull new zealand rabbit or BALb/c mouse for use, after initial immunity uses 200ug (new zealand rabbit) or 20ug (BALb/c mouse) antigen and equal-volume Freund's complete adjuvant (FCA) fully emulsified, the subcutaneous multi-point injection in the back.Behind the initial immunity 21,42,63 days, with Freund's incomplete adjuvant (FIA) emulsive antigen protein fully, each booster immunization 1 time, consumption was the same.Each immunity back 7 ~ 10 days, the ELISA method detects serum titer, reaches 1 * 10 ^-4The time, bloodletting separation of serum .Western blot identifies antibodies specific.

Monoclonal Antibody: immune BALb/c mouse is the same, gets spleen and makes the B cell suspension, with the myeloma cell SP2/0 fusion of logarithmic phase, by HAT (H: xanthoglobulin; A: aminopterin-induced syndrome; T: thymidine) selectivity is cultivated, and obtains hybrid cell line, detects antibody titer by the ELISA method again, filters out specific hybridoma cell line, and obtains monoclonal antibody.

Sequence table

＜110〉Sinogenomax Co., Ltd.

＜120〉polynucleotide of cell death inducing and coded polypeptide thereof and purposes

<130>

<160>26

<170>PatentIn?version?3.3

<210>1

<211>1515

<212>DNA

＜213〉people

<220>

<221>CDS

<222>(217)..(804)

<400>1

cggaggagag?cgcaggagga?aacagtaccg?gctggaggcc?ggtcttgcag?gagcggggga 60

ctgctggggg?cggggcttgg?tggtgactgc?tggcggggcg?gggcctgggg?ctcagagggg 120

tgggctttgg?agatcagagg?gtcgacgctg?cttcgttgcc?tggactctgg?tttccgccct 180

ggagcaagcc?ggggcctggt?cggcagctgg?gccgcc?atg?gag?tcc?acg?ctg?ggc 234

Met?Glu?Ser?Thr?Leu?Gly

1 5

gcg?ggc?atc?gtg?ata?gcc?gag?gcg?cta?cag?aac?cag?cta?gcc?tgg?ctg 282

Ala?Gly?Ile?Val?Ile?Ala?Glu?Ala?Leu?Gln?Asn?Gln?Leu?Ala?Trp?Leu

10 15 20

gag?aac?gtg?tgg?ctc?tgg?atc?acc?ttt?ctg?ggc?gat?ccc?aag?atc?ctc 330

Glu?Asn?Val?Trp?Leu?Trp?Ile?Thr?Phe?Leu?Gly?Asp?Pro?Lys?Ile?Leu

25 30 35

ttt?ctg?ttc?tac?ttc?ccc?gcg?gcc?tac?tac?gcc?tcc?cgc?cgt?gtg?ggc 378

Phe?Leu?Phe?Tyr?Phe?Pro?Ala?Ala?Tyr?Tyr?Ala?Ser?Arg?Arg?Val?Gly

40 45 50

atc?gcg?gtg?ctc?tgg?atc?agc?ctc?atc?acc?gag?tgg?ctc?aac?ctc?atc 426

Ile?Ala?Val?Leu?Trp?Ile?Ser?Leu?Ile?Thr?Glu?Trp?Leu?Asn?Leu?Ile

55 60 65 70

ttc?aag?tgg?ttt?ctt?ttt?gga?gac?agg?ccc?ttt?tgg?tgg?gtc?cat?gag 474

Phe?Lys?Trp?Phe?Leu?Phe?Gly?Asp?Arg?Pro?Phe?Trp?Trp?Val?His?Glu

75 80 85

tct?ggt?tac?tac?agc?cag?gct?cca?gcc?cag?gtt?cac?cag?ttc?ccc?tct 522

Ser?Gly?Tyr?Tyr?Ser?Gln?Ala?Pro?Ala?Gln?Val?His?Gln?Phe?Pro?Ser

90 95 100

tct?tgt?gag?act?ggt?cca?ggc?agc?cct?tct?gga?cac?tgc?atg?atc?aca 570

Ser?Cys?Glu?Thr?Gly?Pro?Gly?Ser?Pro?Ser?Gly?His?Cys?Met?Ile?Thr

105 110 115

gga?gca?gcc?ctc?tgg?ccc?ata?atg?acg?gcc?ctg?tct?tcg?cag?gtg?gcc 618

Gly?Ala?Ala?Leu?Trp?Pro?Ile?Met?Thr?Ala?Leu?Ser?Ser?Gln?Val?Ala

120 125 130

act?cgg?gcc?cgc?agg?cgc?tgt?cct?ggg?ctg?gct?gat?gac?tcc?ccg?agt 666

Thr?Arg?Ala?Arg?Arg?Arg?Cys?Pro?Gly?Leu?Ala?Asp?Asp?Ser?Pro?Ser

135 140 145 150

gcc?tat?gga?gcg?gga?gct?aag?ctt?cta?tgg?gtt?gac?tgc?act?ggc?cct 714

Ala?Tyr?Gly?Ala?Gly?Ala?Lys?Leu?Leu?Trp?Val?Asp?Cys?Thr?Gly?Pro

155 160 165

cat?gct?agg?cac?cag?cct?cat?cta?ttg?gac?cct?ctt?tac?act?ggg?cct 762

His?Ala?Arg?His?Gln?Pro?His?Leu?Leu?Asp?Pro?Leu?Tyr?Thr?Gly?Pro

170 175 180

gga?tct?ttc?ttg?gtc?cat?cag?cct?agc?ctt?caa?gtg?gtg?tga 804

Gly?Ser?Phe?Leu?Val?His?Gln?Pro?Ser?Leu?Gln?Val?Val

185 190 195

gcggcctgag?tggatacacg?tggatagccg?gccctttgcc?tccctgagcc?gtgactcagg 864

ggctgccctg?ggcctgggca?ttgccttgca?ctctccctgc?tatgcccagg?tgcgtcgggc 924

acagctggga?aatggccaga?agatagcctg?ccttgtgctg?gccatggggc?tgctgggccc 984

cctggactgg?ctgggccacc?cccctcagat?cagcctcttc?tacattttca?atttcctcaa 1044

gtacaccctc?tggccatgcc?tagtcctggc?cctcgtgccc?tgggcagtgc?acatgttcag 1104

tgcccaggaa?gcaccgccca?tccactcttc?ctgacttctt?gtgtgcctcc?ctttcctttc 1164

cctcccacaa?agccaacact?ctgtgaccac?cacactccag?gaggcagccc?catccccttc 1224

cagcccctaa?gtaggccctc?ccctccctaa?atctgcttcc?gcaccacctg?gtcttagccc 1284

caaagatggg?ccttctctct?cccagataag?ttggtcctcc?ctctgccttt?cctctcaagc 1344

ccccaaagag?caaaggcaac?agcaagacca?gcgggttctt?gcaacactgt?gaggggcagc 1404

cagggcggcc?ccaataaagc?ccttgaatac?tttgaaaaaa?aaaaaaaaaa?aaaaaaaaaa 1464

aaaaaaaaaa?aaaaaaaaaa?aaaaaaaaaa?aaaaaaaaaa?aaaaaaaaaa?a 1515

<210>2

<211>195

<212>PRT

＜213〉people

<400>2

Met?Glu?Ser?Thr?Leu?Gly?Ala?Gly?Ile?Val?Ile?Ala?Glu?Ala?Leu?Gln

1 5 10 15

Asn?Gln?Leu?Ala?Trp?Leu?Glu?Asn?Val?Trp?Leu?Trp?Ile?Thr?Phe?Leu

20 25 30

Gly?Asp?Pro?Lys?Ile?Leu?Phe?Leu?Phe?Tyr?Phe?Pro?Ala?Ala?Tyr?Tyr

35 40 45

Ala?Ser?Arg?Arg?Val?Gly?Ile?Ala?Val?Leu?Trp?Ile?Ser?Leu?Ile?Thr

50 55 60

Glu?Trp?Leu?Asn?Leu?Ile?Phe?Lys?Trp?Phe?Leu?Phe?Gly?Asp?Arg?Pro

65 70 75 80

Phe?Trp?Trp?Val?His?Glu?Ser?Gly?Tyr?Tyr?Ser?Gln?Ala?Pro?Ala?Gln

85 90 95

Val?His?Gln?Phe?Pro?Ser?Ser?Cys?Glu?Thr?Gly?Pro?Gly?Ser?Pro?Ser

100 105 110

Gly?His?Cys?Met?Ile?Thr?Gly?Ala?Ala?Leu?Trp?Pro?Ile?Met?Thr?Ala

115 120 125

Leu?Ser?Ser?Gln?Val?Ala?Thr?Arg?Ala?Arg?Arg?Arg?Cys?Pro?Gly?Leu

130 135 140

Ala?Asp?Asp?Ser?Pro?Ser?Ala?Tyr?Gly?Ala?Gly?Ala?Lys?Leu?Leu?Trp

145 150 155 160

Val?Asp?Cys?Thr?Gly?Pro?His?Ala?Arg?His?Gln?Pro?His?Leu?Leu?Asp

165 170 175

Pro?Leu?Tyr?Thr?Gly?Pro?Gly?Ser?Phe?Leu?Val?His?Gln?Pro?Ser?Leu

180 185 190

Gln?Val?Val

195

<210>3

<211>2251

<212>DNA

＜213〉people

<220>

<221>CDS

<222>(186)..(1598)

<400>3

gacgtcacgg?tcactgacag?cgtgagcccg?cggcggctgc?tgccatggtg?gctggcggcc 60

gggtaagggt?ctgagtggat?ctcctgccag?gccagagcgc?cttcgggggc?cgcggcggaa 120

ggccaggagt?ttgcagccag?ggcgccgggt?ttgtggtctg?cagtgtcgtg?aggctgaggt 180

gcagc?atg?tct?aga?ctg?gga?gcc?ctg?ggt?ggt?gcc?cgt?gcc?ggg?ctg?gga 230

Met?Ser?Arg?Leu?Gly?Ala?Leu?Gly?Gly?Ala?Arg?Ala?Gly?Leu?Gly

1 5 10 15

ctg?ttg?ctg?ggt?acc?gcc?gcc?ggc?ctt?gga?ttc?ctg?tgc?ctc?ctt?tac 278

Leu?Leu?Leu?Gly?Thr?Ala?Ala?Gly?Leu?Gly?Phe?Leu?Cys?Leu?Leu?Tyr

20 25 30

agc?cag?cga?tgg?aaa?cgg?acc?cag?cgt?cat?ggc?cgc?agc?cag?agc?ctg 326

Ser?Gln?Arg?Trp?Lys?Arg?Thr?Gln?Arg?His?Gly?Arg?Ser?Gln?Ser?Leu

35 40 45

ccc?aac?tcc?ctg?gac?tat?acg?cag?act?tca?gat?ccc?gga?cgc?cac?gtg 374

Pro?Asn?Ser?Leu?Asp?Tyr?Thr?Gln?Thr?Ser?Asp?Pro?Gly?Arg?His?Val

50 55 60

atg?ctc?ctg?cgg?gct?gtc?cca?ggt?ggg?gct?gga?gat?gcc?tca?gtg?ctg 422

Met?Leu?Leu?Arg?Ala?Val?Pro?Gly?Gly?Ala?Gly?Asp?Ala?Ser?Val?Leu

65 70 75

ccc?agc?ctt?cca?cgg?gaa?gga?cag?gag?aag?gtg?ctg?gac?cgc?ctg?gac 470

Pro?Ser?Leu?Pro?Arg?Glu?Gly?Gln?Glu?Lys?Val?Leu?Asp?Arg?Leu?Asp

80 85 90 95

ttt?gtg?ctg?acc?agc?ctt?gtg?gcg?ctg?cgg?cgg?gag?gtg?gag?gag?ctg 518

Phe?Val?Leu?Thr?Ser?Leu?Val?Ala?Leu?Arg?Arg?Glu?Val?Glu?Glu?Leu

100 105 110

aga?agc?agc?ctg?cga?ggg?ctt?gcg?ggg?gag?att?gtt?ggg?gag?gtc?cga 566

Arg?Ser?Ser?Leu?Arg?Gly?Leu?Ala?Gly?Glu?Ile?Val?Gly?Glu?Val?Arg

115 120 125

tgc?cac?atg?gaa?gag?aac?cag?aga?gtg?gct?cgg?cgg?cga?agg?ttt?ccg 614

Cys?His?Met?Glu?Glu?Asn?Gln?Arg?Val?Ala?Arg?Arg?Arg?Arg?Phe?Pro

130 135 140

ttt?gtc?cgg?gag?agg?agt?gac?tcc?act?ggc?tcc?agc?tct?gtc?tac?ttc 662

Phe?Val?Arg?Glu?Arg?Ser?Asp?Ser?Thr?Gly?Ser?Ser?Ser?Val?Tyr?Phe

145 150 155

acg?gcc?tcc?tcg?gga?gcc?acg?ttc?aca?gat?gct?gag?agt?gaa?ggg?ggt 710

Thr?Ala?Ser?Ser?Gly?Ala?Thr?Phe?Thr?Asp?Ala?Glu?Ser?Glu?Gly?Gly

160 165 170 175

tac?aca?aca?gcc?aat?gcg?gag?tct?gac?aat?gag?cgg?gac?tct?gac?aaa 758

Tyr?Thr?Thr?Ala?Asn?Ala?Glu?Ser?Asp?Asn?Glu?Arg?Asp?Ser?Asp?Lys

180 185 190

gaa?agt?gag?gac?ggg?gaa?gat?gaa?gtg?agc?tgt?gag?act?gtg?aag?atg 806

Glu?Ser?Glu?Asp?Gly?Glu?Asp?Glu?Val?Ser?Cys?Glu?Thr?Val?Lys?Met

195 200 205

ggg?aga?aag?gat?tct?ctt?gac?ttg?gag?gaa?gag?gca?gct?tca?ggt?gcc 854

Gly?Arg?Lys?Asp?Ser?Leu?Asp?Leu?Glu?Glu?Glu?Ala?Ala?Ser?Gly?Ala

210 215 220

tcc?agt?gcc?ctg?gag?gct?gga?ggt?tcc?tca?ggc?ttg?gag?gat?gtg?ctg 902

Ser?Ser?Ala?Leu?Glu?Ala?Gly?Gly?Ser?Ser?Gly?Leu?Glu?Asp?Val?Leu

225 230 235

ccc?ctc?ctg?cag?cag?gcc?gac?gag?ctg?cac?agg?ggt?gat?gag?caa?ggc 950

Pro?Leu?Leu?Gln?Gln?Ala?Asp?Glu?Leu?His?Arg?Gly?Asp?Glu?Gln?Gly

240 245 250 255

aag?cgg?gag?ggc?ttc?cag?ctg?ctg?ctc?aac?aac?aag?ctg?gtg?tat?gga 998

Lys?Arg?Glu?Gly?Phe?Gln?Leu?Leu?Leu?Asn?Asn?Lys?Leu?Val?Tyr?Gly

260 265 270

agc?cgg?cag?gac?ttt?ctc?tgg?cgc?ctg?gcc?cga?gcc?tac?agt?gac?atg 1046

Ser?Arg?Gln?Asp?Phe?Leu?Trp?Arg?Leu?Ala?Arg?Ala?Tyr?Ser?Asp?Met

275 280 285

tgt?gag?ctc?act?gag?gag?gtg?agc?gag?aag?aag?tca?tat?gcc?cta?gat 1094

Cys?Glu?Leu?Thr?Glu?Glu?Val?Ser?Glu?Lys?Lys?Ser?Tyr?Ala?Leu?Asp

290 295 300

gga?aaa?gaa?gaa?gca?gag?gct?gct?ctg?gag?aag?ggg?gat?gag?agt?gct 1142

Gly?Lys?Glu?Glu?Ala?Glu?Ala?Ala?Leu?Glu?Lys?Gly?Asp?Glu?Ser?Ala

305 310 315

gac?tgt?cac?ctg?tgg?tat?gcg?gtg?ctt?tgt?ggt?cag?ctg?gct?gag?cat 1190

Asp?Cys?His?Leu?Trp?Tyr?Ala?Val?Leu?Cys?Gly?Gln?Leu?Ala?Glu?His

320 325 330 335

gag?agc?atc?cag?agg?cgc?atc?cag?agt?ggc?ttt?agc?ttc?aag?gag?cat 1238

Glu?Ser?Ile?Gln?Arg?Arg?Ile?Gln?Ser?Gly?Phe?Ser?Phe?Lys?Glu?His

340 345 350

gtg?gac?aaa?gcc?att?gct?ctc?cag?cca?gaa?aac?ccc?atg?gct?cac?ttt 1286

Val?Asp?Lys?Ala?Ile?Ala?Leu?Gln?Pro?Glu?Asn?Pro?Met?Ala?His?Phe

355 360 365

ctt?ctt?ggc?agg?tgg?tgc?tat?cag?gtc?tct?cac?ctg?agc?tgg?cta?gaa 1334

Leu?Leu?Gly?Arg?Trp?Cys?Tyr?Gln?Val?Ser?His?Leu?Ser?Trp?Leu?Glu

370 375 380

aaa?aaa?act?gct?aca?gcc?ttg?ctt?gaa?agc?cct?ctc?agt?gcc?act?gtg 1382

Lys?Lys?Thr?Ala?Thr?Ala?Leu?Leu?Glu?Ser?Pro?Leu?Ser?Ala?Thr?Val

385 390 395

gaa?gat?gcc?ctc?cag?agc?ttc?cta?aag?gct?gaa?gaa?cta?cag?cca?gga 1430

Glu?Asp?Ala?Leu?Gln?Ser?Phe?Leu?Lys?Ala?Glu?Glu?Leu?Gln?Pro?Gly

400 405 410 415

ttt?tcc?aaa?gca?gga?agg?gta?tat?att?tcc?aag?tgc?tac?aga?gaa?cta 1478

Phe?Ser?Lys?Ala?Gly?Arg?Val?Tyr?Ile?Ser?Lys?Cys?Tyr?Arg?Glu?Leu

420 425 430

ggg?aaa?aac?tct?gaa?gct?aga?tgg?tgg?atg?aag?ttg?gcc?ctg?gag?ctg 1526

Gly?Lys?Asn?Ser?Glu?Ala?Arg?Trp?Trp?Met?Lys?Leu?Ala?Leu?Glu?Leu

435 440 445

cca?gat?gtc?acg?aag?gag?gat?ttg?gct?atc?cag?aag?gac?ctg?gaa?gaa 1574

Pro?Asp?Val?Thr?Lys?Glu?Asp?Leu?Ala?Ile?Gln?Lys?Asp?Leu?Glu?Glu

450 455 460

ctg?gaa?gtc?att?tta?cga?gac?taa?ccacgtttca?ctggccttca?tgacttgatg 1628

Leu?Glu?Val?Ile?Leu?Arg?Asp

465 470

ccactattta?aggtgggggg?gcggggaggc?ttttttcctt?agaccttgct?gagatcagga 1688

aaccacacaa?atctgtctcc?tgggtctgac?tgctacccac?taccactccc?cattagttaa 1748

tttattctaa?cctctaacct?aatctagaat?tggggcagta?ctcatggctt?ccgtttctgt 1808

tgttctctcc?cttgagtaat?ctcttaaaaa?aatcaagatt?cacacctgcc?ccaggattac 1868

acatgggtag?agcctgcaag?acctgagacc?ttccaattgc?tggtgaggtg?gatgaacttc 1928

aaagctatag?gaacaaagca?cataacttgt?cactttaatc?tttttcactg?actaatagga 1988

ctcagtacat?atagtcttaa?gatcatacct?tacctaccaa?ggtaaaaaga?gggatcagag 2048

tggcccacag?acattgcttt?cttatcacct?atcatgtgaa?ttctacctgt?attccagggc 2108

tggaccactt?gataacttcc?agtgtcctgg?cagcttttgg?aatgacagca?gtggtatggg 2168

gtttatgatg?ctataaaaca?atgtctgaaa?agttgcctag?aatatatttt?gttacaaact 2228

tgaaataaac?caaatttgat?gtt 2251

<210>4

<211>470

<212>PRT

＜213〉people

<400>4

Met?Ser?Arg?Leu?Gly?Ala?Leu?Gly?Gly?Ala?Arg?Ala?Gly?Leu?Gly?Leu

1 5 10 15

Leu?Leu?Gly?Thr?Ala?Ala?Gly?Leu?Gly?Phe?Leu?Cys?Leu?Leu?Tyr?Ser

20 25 30

Gln?Arg?Trp?Lys?Arg?Thr?Gln?Arg?His?Gly?Arg?Ser?Gln?Ser?Leu?Pro

35 40 45

Asn?Ser?Leu?Asp?Tyr?Thr?Gln?Thr?Ser?Asp?Pro?Gly?Arg?His?Val?Met

50 55 60

Leu?Leu?Arg?Ala?Val?Pro?Gly?Gly?Ala?Gly?Asp?Ala?Ser?Val?Leu?Pro

65 70 75 80

Ser?Leu?Pro?Arg?Glu?Gly?Gln?Glu?Lys?Val?Leu?Asp?Arg?Leu?Asp?Phe

85 90 95

Val?Leu?Thr?Ser?Leu?Val?Ala?Leu?Arg?Arg?Glu?Val?Glu?Glu?Leu?Arg

100 105 110

Ser?Ser?Leu?Arg?Gly?Leu?Ala?Gly?Glu?Ile?Val?Gly?Glu?Val?Arg?Cys

115 120 125

His?Met?Glu?Glu?Asn?Gln?Arg?Val?Ala?Arg?Arg?Arg?Arg?Phe?Pro?Phe

130 135 140

Val?Arg?Glu?Arg?Ser?Asp?Ser?Thr?Gly?Ser?Ser?Ser?Val?Tyr?Phe?Thr

145 150 155 160

Ala?Ser?Ser?Gly?Ala?Thr?Phe?Thr?Asp?Ala?Glu?Ser?Glu?Gly?Gly?Tyr

165 170 175

Thr?Thr?Ala?Asn?Ala?Glu?Ser?Asp?Asn?Glu?Arg?Asp?Ser?Asp?Lys?Glu

180 185 190

Ser?Glu?Asp?Gly?Glu?Asp?Glu?Val?Ser?Cys?Glu?Thr?Val?Lys?Met?Gly

195 200 205

Arg?Lys?Asp?Ser?Leu?Asp?Leu?Glu?Glu?Glu?Ala?Ala?Ser?Gly?Ala?Ser

210 215 220

Ser?Ala?Leu?Glu?Ala?Gly?Gly?Ser?Ser?Gly?Leu?Glu?Asp?Val?Leu?Pro

225 230 235 240

Leu?Leu?Gln?Gln?Ala?Asp?Glu?Leu?His?Arg?Gly?Asp?Glu?Gln?Gly?Lys

245 250 255

Arg?Glu?Gly?Phe?Gln?Leu?Leu?Leu?Asn?Asn?Lys?Leu?Val?Tyr?Gly?Ser

260 265 270

Arg?Gln?Asp?Phe?Leu?Trp?Arg?Leu?Ala?Arg?Ala?Tyr?Ser?Asp?Met?Cys

275 280 285

Glu?Leu?Thr?Glu?Glu?Val?Ser?Glu?Lys?Lys?Ser?Tyr?Ala?Leu?Asp?Gly

290 295 300

Lys?Glu?Glu?Ala?Glu?Ala?Ala?Leu?Glu?Lys?Gly?Asp?Glu?Ser?Ala?Asp

305 310 315 320

Cys?His?Leu?Trp?Tyr?Ala?Val?Leu?Cys?Gly?Gln?Leu?Ala?Glu?His?Glu

325 330 335

Ser?Ile?Gln?Arg?Arg?Ile?Gln?Ser?Gly?Phe?Ser?Phe?Lys?Glu?His?Val

340 345 350

Asp?Lys?Ala?Ile?Ala?Leu?Gln?Pro?Glu?Asn?Pro?Met?Ala?His?Phe?Leu

355 360 365

Leu?Gly?Arg?Trp?Cys?Tyr?Gln?Val?Ser?His?Leu?Ser?Trp?Leu?Glu?Lys

370 375 380

Lys?Thr?Ala?Thr?Ala?Leu?Leu?Glu?Ser?Pro?Leu?Ser?Ala?Thr?Val?Glu

385 390 395 400

Asp?Ala?Leu?Gln?Ser?Phe?Leu?Lys?Ala?Glu?Glu?Leu?Gln?Pro?Gly?Phe

405 410 415

Ser?Lys?Ala?Gly?Arg?Val?Tyr?Ile?Ser?Lys?Cys?Tyr?Arg?Glu?Leu?Gly

420 425 430

Lys?Asn?Ser?Glu?Ala?Arg?Trp?Trp?Met?Lys?Leu?Ala?Leu?Glu?Leu?Pro

435 440 445

Asp?Val?Thr?Lys?6lu?Asp?Leu?Ala?Ile?Gln?Lys?Asp?Leu?Glu?Glu?Leu

450 455 460

Glu?Val?Ile?Leu?Arg?Asp

465 470

<210>5

<211>2310

<212>DNA

＜213〉people

<220>

<221>CDS

<222>(148)..(675)

<400>5

cggacgcgtg?gggagaggct?gtttaccaga?acagcataac?aagggcaggt?ctgactgcaa 60

ggctgggact?gggaggcaga?gccgccgcca?agggggcctc?ggttaaacac?tggtcgttca 120

atcacctgca?agacgaagga?ggcaagg?atg?ctg?ttg?gcc?tgg?gta?caa?gca?ttc 174

Met?Leu?Leu?Ala?Trp?Val?Gln?Ala?Phe

1 5

ctc?gtc?agc?aac?atg?ctc?cta?gca?gaa?gcc?tat?gga?tct?gga?ggg?gcc 222

Leu?Val?Ser?Asn?Met?Leu?Leu?Ala?Glu?Ala?Tyr?Gly?Ser?Gly?Gly?Ala

10 15 20 25

ggc?aat?cac?agt?tac?tgc?cga?aac?ccg?gac?gag?gac?ccg?cgc?ggg?ccc 270

Gly?Asn?His?Ser?Tyr?Cys?Arg?Asn?Pro?Asp?Glu?Asp?Pro?Arg?Gly?Pro

30 35 40

tgg?tgc?tac?gtc?agt?ggc?gag?gcc?ggc?gtc?cct?gag?aaa?cgg?cct?tgc 318

Trp?Cys?Tyr?Val?Ser?Gly?Glu?Ala?Gly?Val?Pro?Glu?Lys?Arg?Pro?Cys

45 50 55

gag?gac?ctg?cgc?tgt?cca?gag?acc?acc?tcc?cag?gcc?ctg?cca?gcc?ttc 366

Glu?Asp?Leu?Arg?Cys?Pro?Glu?Thr?Thr?Ser?Gln?Ala?Leu?Pro?Ala?Phe

60 65 70

acg?aca?gaa?atc?cag?gaa?gcg?tct?gaa?ggg?cca?ggt?gca?gat?gag?gtg 414

Thr?Thr?Glu?Ile?Gln?Glu?Ala?Ser?Glu?Gly?Pro?Gly?Ala?Asp?Glu?Val

75 80 85

cag?gtg?ttc?gct?cct?gcc?aac?gcc?ctg?ccc?gct?cgg?agt?gag?gcg?gca 462

Gln?Val?Phe?Ala?Pro?Ala?Asn?Ala?Leu?Pro?Ala?Arg?Ser?Glu?Ala?Ala

90 95 100 105

gct?gtg?cag?cca?gtg?att?ggg?atc?agc?cag?cgg?gtg?cgg?atg?aac?tcc 510

Ala?Val?Gln?Pro?Val?Ile?Gly?Ile?Ser?Gln?Arg?Val?Arg?Met?Asn?Ser

110 115 120

aag?gag?aaa?aag?gac?ctg?gga?act?ctg?ggc?tac?gtg?ctg?ggc?att?acc 558

Lys?Glu?Lys?Lys?Asp?Leu?Gly?Thr?Leu?Gly?Tyr?Val?Leu?Gly?Ile?Thr

125 130 135

atg?atg?gtg?atc?atc?att?gcc?atc?gga?gct?ggc?atc?atc?ttg?ggc?tac 606

Met?Met?Val?Ile?Ile?Ile?Ala?Ile?Gly?Ala?Gly?Ile?Ile?Leu?Gly?Tyr

140 145 150

tcc?tac?aag?agg?ggg?aag?gat?ttg?aaa?gaa?cag?cat?gat?cag?aaa?gta 654

Ser?Tyr?Lys?Arg?Gly?Lys?Asp?Leu?Lys?Glu?Gln?His?Asp?Gln?Lys?Val

155 160 165

tgt?gag?agg?gag?atg?cag?tga?atcactctgc?ccttgtctgc?cttcaccaac 705

Cys?Glu?Arg?Glu?Met?Gln

170 175

cccacctgtg?agattgtgga?tgagaagact?gtcgtggtcc?acaccagcca?gactccagtt 765

gaccctcagg?agggcagcac?cccccttatg?ggccaggccg?ggactcctgg?ggcctgagcc 825

cccccagtgg?gcaggagccc?atgcagacac?tggtgcagga?cagcccaccc?tcctacagct 885

aggaggaact?accactttgt?gttctggtta?aaaccctacc?actcccccgc?ttttttggcg 945

aatcctagta?agagtgacag?aagcaggtgg?ccctgtgggc?tgagggtaag?gctgggtagg 1005

gtcctaacag?tgctccttgt?ccatcccttg?gagcagattt?tgtctgtgga?tggagacagt 1065

ggcagctccc?acagtgatgc?tgctgctaag?ggcttccaaa?cattgcctgc?acccctggaa 1125

ctgaaccagg?gatagacggg?gagctccccc?aggctcctct?gtgctttact?aagatggcct 1185

cagtctccac?tgtgggcttg?agtggcatac?actgttattc?atggttaagg?taaagcaggt 1245

caagggatgg?cattgaaaaa?atatatttag?tttttaaaat?atttgggatg?gaactcccta 1305

ctgacctctg?agaactggaa?acgagtttgt?acagaagtca?gaactttggg?ttgggaatga 1365

gatctaggtt?gtggctgctg?gtatgcttca?gcttgctggc?aatgatgtgc?cttgacaacc 1425

gtgggccagg?cctgggccca?gggactcttc?ctgtttcata?aggaaaggaa?gaattgcact 1485

gagcattcca?cttaggaaga?ggatagagaa?ggatctgctc?cgcctttggc?cacaggagca 1545

gaggcagacc?tgggatgccc?cagtttctct?tcagggatgg?atagtgacct?gtcttcattt 1605

tgcacaggta?agagagtagt?tagctaacct?atgggaatta?tactgtgggg?ccttgtgagc 1665

tgcttctaag?aggctaacct?ggaaactaag?ctcagaggca?aggtaataaa?gcacttcagg 1725

gcttgctccc?caagtgggcc?tgatttagca?ggtggtcctg?cgggcgtcca?ggtcagcacc 1785

ttcctgtagg?gcactggggc?tagggtcaca?gcccctaact?cataaagcaa?tcaaagaacc 1845

attagaaagg?gctcattaag?ccttttggac?acaggacccc?agagaggaaa?aagtgacttg 1905

cccaaggtcg?taagcaagct?actggcatgg?caagagccca?gcttcctgac?ggagcgcaac 1965

atttctccac?tgcactgtgc?tagcagctca?gcagggcctc?taacctgtga?tgtcacactc 2025

aagaggcctt?ggcagctcct?agccatagag?cttcctttcc?agaacccttc?cactgcccaa 2085

tgtggagaca?ggggttagtg?gggctttcta?tggagccatc?tgctttgggg?acctagacct 2145

caggtggtct?cttggtgtta?gtgatgctgg?agaagagaat?attactggtt?tctacttttc 2205

tataaaggca?tttctctata?tacatgtttt?atatacctca?ttctgacacc?tgcatatagt 2265

gtgggaaatt?gctctgcatt?tgacttaatt?aaaaaaaaaa?aaaaa 2310

<210>6

<211>175

<212>PRT

＜213〉people

<400>6

Met?Leu?Leu?Ala?Trp?Val?Gln?Ala?Phe?Leu?Val?Ser?Asn?Met?Leu?Leu

1 5 10 15

Ala?Glu?Ala?Tyr?Gly?Ser?Gly?Gly?Ala?Gly?Asn?His?Ser?Tyr?Cys?Arg

20 25 30

Asn?Pro?Asp?Glu?Asp?Pro?Arg?Gly?Pro?Trp?Cys?Tyr?Val?Ser?Gly?Glu

35 40 45

Ala?Gly?Val?Pro?Glu?Lys?Arg?Pro?Cys?Glu?Asp?Leu?Arg?Cys?Pro?Glu

50 55 60

Thr?Thr?Ser?Gln?Ala?Leu?Pro?Ala?Phe?Thr?Thr?Glu?Ile?Gln?Glu?Ala

65 70 75 80

Ser?Glu?Gly?Pro?Gly?Ala?Asp?Glu?Val?Gln?Val?Phe?Ala?Pro?Ala?Asn

85 90 95

Ala?Leu?Pro?Ala?Arg?Ser?Glu?Ala?Ala?Ala?Val?Gln?Pro?Val?Ile?Gly

100 105 110

Ile?Ser?Gln?Arg?Val?Arg?Met?Asn?Ser?Lys?Glu?Lys?Lys?Asp?Leu?Gly

115 120 125

Thr?Leu?Gly?Tyr?Val?Leu?Gly?Ile?Thr?Met?Met?Val?Ile?Ile?Ile?Ala

130 135 140

Ile?Gly?Ala?Gly?Ile?Ile?Leu?Gly?Tyr?Ser?Tyr?Lys?Arg?Gly?Lys?Asp

145 150 155 160

Leu?Lys?Glu?Gln?His?Asp?Gln?Lys?Val?Cys?Glu?Arg?Glu?Met?Gln

165 170 175

<210>7

<211>846

<212>DNA

＜213〉people

<220>

<221>CDS

<222>(102)..(653)

<400>7

ggcacgaggg?agcacggccc?gcgggcggcg?ttcgctggag?ctggtggacc?gggcggcggg 60

gcagaccgct?ggggactgcg?ggcggcgctg?tgtccgtcgc?c?atg?aca?gat?cag?acc 116

Met?Thr?Asp?Gln?Thr

1 5

tat?tgt?gac?cgc?ctg?gtg?cag?gac?acg?cct?ttc?ctg?aca?ggc?cat?ggg 164

Tyr?Cys?Asp?Arg?Leu?Val?Gln?Asp?Thr?Pro?Phe?Leu?Thr?Gly?His?Gly

10 15 20

cgc?ttg?agt?gag?cag?cag?gtg?gac?agg?atc?atc?ctc?cag?ctg?aac?cgt 212

Arg?Leu?Ser?Glu?Gln?Gln?Val?Asp?Arg?Ile?Ile?Leu?Gln?Leu?Asn?Arg

25 30 35

tac?tac?cca?cag?atc?ctt?acc?aac?aag?gag?gcg?gaa?aag?ttc?cgg?aac 260

Tyr?Tyr?Pro?Gln?Ile?Leu?Thr?Asn?Lys?Glu?Ala?Glu?Lys?Phe?Arg?Asn

40 45 50

ccc?aag?gca?tcc?ttg?cgt?gtg?cgg?ctc?tgt?gac?ctc?ctg?agc?cac?ctg 308

Pro?Lys?Ala?Ser?Leu?Arg?Val?Arg?Leu?Cys?Asp?Leu?Leu?Ser?His?Leu

55 60 65

cag?cgg?agc?ggt?gag?cgg?gac?tgc?cag?gag?ttc?tac?cga?gcc?ctg?tat 356

Gln?Arg?Ser?Gly?Glu?Arg?Asp?Cys?Gln?Glu?Phe?Tyr?Arg?Ala?Leu?Tyr

70 75 80 85

atc?cat?gcc?cag?ccc?ctg?cac?agc?cgc?ctg?ccc?agc?cgc?cac?gct?ctg 404

Ile?His?Ala?Gln?Pro?Leu?His?Ser?Arg?Leu?Pro?Ser?Arg?His?Ala?Leu

90 95 100

cag?aac?tca?gat?tgc?aca?gag?cta?gac?tcg?ggc?agc?cag?agc?ggc?gag 452

Gln?Asn?Ser?Asp?Cys?Thr?Glu?Leu?Asp?Ser?Gly?Ser?Gln?Ser?Gly?Glu

105 110 115

ctg?agt?aac?agg?gga?ccc?atg?agc?ttc?ctg?gct?ggc?ctg?ggc?ctt?gct 500

Leu?Ser?Asn?Arg?Gly?Pro?Met?Ser?Phe?Leu?Ala?Gly?Leu?Gly?Leu?Ala

120 125 130

gtg?gga?ctg?gcc?ctg?ctc?ctg?tac?tgc?tat?ccg?cca?gac?ccc?aag?ggc 548

Val?Gly?Leu?Ala?Leu?Leu?Leu?Tyr?Cys?Tyr?Pro?Pro?Asp?Pro?Lys?Gly

135 140 145

ctg?cca?ggg?acc?cgg?cgc?gtc?ctc?ggt?ttc?tcg?cct?gtc?atc?atc?gac 596

Leu?Pro?Gly?Thr?Arg?Arg?Val?Leu?Gly?Phe?Ser?Pro?Val?Ile?Ile?Asp

150 155 160 165

aga?cat?gtc?agc?cgc?tac?ctg?ctg?gcc?ttc?ctg?gca?gat?gac?cta?ggg 644

Arg?His?Val?Ser?Arg?Tyr?Leu?Leu?Ala?Phe?Leu?Ala?Asp?Asp?Leu?Gly

170 175 180

ggg?ctc?tga?cagaccctgg?acccagggcc?tcacctgcca?ctcaaccaaa 693

Gly?Leu

gagtcctcga?gccggcctgc?caaggggact?gctgcttctt?tttctaaatg?catatttttc 753

attatttata?atttgtgtaa?aaaacacacc?ttcaccttac?aaggtgctga?ccatattaaa 813

tgttcaagtt?ctctcaaaaa?aaaaaaaaaa?aaa 846

<210>8

<211>183

<212>PRT

＜213〉people

<400>8

Met?Thr?Asp?Gln?Thr?Tyr?Cys?Asp?Arg?Leu?Val?Gln?Asp?Thr?Pro?Phe

1 5 10 15

Leu?Thr?Gly?His?Gly?Arg?Leu?Ser?Glu?Gln?Gln?Val?Asp?Arg?Ile?Ile

20 25 30

Leu?Gln?Leu?Asn?Arg?Tyr?Tyr?Pro?Gln?Ile?Leu?Thr?Asn?Lys?Glu?Ala

35 40 45

Glu?Lys?Phe?Arg?Asn?Pro?Lys?Ala?Ser?Leu?Arg?Val?Arg?Leu?Cys?Asp

50 55 60

Leu?Leu?Ser?His?Leu?Gln?Arg?Ser?Gly?Glu?Arg?Asp?Cys?Gln?Glu?Phe

65 70 75 80

Tyr?Arg?Ala?Leu?Tyr?Ile?His?Ala?Gln?Pro?Leu?His?Ser?Arg?Leu?Pro

85 90 95

Ser?Arg?His?Ala?Leu?Gln?Asn?Ser?Asp?Cys?Thr?Glu?Leu?Asp?Ser?Gly

100 105 110

Ser?Gln?Ser?Gly?Glu?Leu?Ser?Asn?Arg?Gly?Pro?Met?Ser?Phe?Leu?Ala

115 120 125

Gly?Leu?Gly?Leu?Ala?Val?Gly?Leu?Ala?Leu?Leu?Leu?Tyr?Cys?Tyr?Pro

130 135 140

Pro?Asp?Pro?Lys?Gly?Leu?Pro?Gly?Thr?Arg?Arg?Val?Leu?Gly?Phe?Ser

145 150 155 160

Pro?Val?Ile?Ile?Asp?Arg?His?Val?Ser?Arg?Tyr?Leu?Leu?Ala?Phe?Leu

165 170 175

Ala?Asp?Asp?Leu?Gly?Gly?Leu

180

<210>9

<211>1753

<212>DNA

＜213〉people

<220>

<221>CDS

<222>(211)..(786)

<400>9

gtttaggggt?gggatcgggt?cgggatccgg?accagccaag?gaagggtcca?ggtgccgcgg 60

atcttgagaa?ggaggcgtgg?cctggcgctg?ggattggcgc?cggatgcacc?ccgggttgtg 120

gagtttgggg?atcctcatcc?ggcgccctct?ccccttccag?gagtgcacag?tctgcgggtg 180

gaccggggaa?agaagacgta?cttcatcgtc?atg?cag?agc?gtc?ttc?tac?ccc?gcc 234

Met?Gln?Ser?Val?Phe?Tyr?Pro?Ala

1 5

ggc?cgc?atc?tcc?gag?agg?tat?gac?atc?aaa?ggc?tgc?gag?gtg?agc?cgc 282

Gly?Arg?Ile?Ser?Glu?Arg?Tyr?Asp?Ile?Lys?Gly?Cys?Glu?Val?Ser?Arg

10 15 20

tgg?gtg?gat?ccc?gcc?cct?gag?ggc?agc?ccc?ctt?gtt?ctg?gtg?ctg?aag 330

Trp?Val?Asp?Pro?Ala?Pro?Glu?Gly?Ser?Pro?Leu?Val?Leu?Val?Leu?Lys

25 30 35 40

gac?ctc?aac?ttt?cag?ggc?aag?acc?atc?aac?ctg?ggg?ccc?cag?cgg?agc 378

Asp?Leu?Asn?Phe?Gln?Gly?Lys?Thr?Ile?Asn?Leu?Gly?Pro?Gln?Arg?Ser

45 50 55

tgg?ttc?ctc?cgc?cag?atg?gaa?ctg?gat?acc?acc?ttc?ctc?cgg?gag?ctc 426

Trp?Phe?Leu?Arg?Gln?Met?Glu?Leu?Asp?Thr?Thr?Phe?Leu?Arg?Glu?Leu

60 65 70

aac?gtg?ctg?gat?tac?agc?ctc?ctg?ata?gcc?ttc?caa?cgt?ctc?cac?gag 474

Asn?Val?Leu?Asp?Tyr?Ser?Leu?Leu?Ile?Ala?Phe?Gln?Arg?Leu?His?Glu

75 80 85

gat?gag?agg?ggc?ccg?ggc?agc?agc?ctc?atc?ttc?cgc?acg?gcc?agg?tct 522

Asp?Glu?Arg?Gly?Pro?Gly?Ser?Ser?Leu?Ile?Phe?Arg?Thr?Ala?Arg?Ser

90 95 100

gtg?caa?ggg?gca?cag?agc?ccg?gaa?gag?tcg?aga?gcc?caa?aac?cgc?cgg 570

Val?Gln?Gly?Ala?Gln?Ser?Pro?Glu?Glu?Ser?Arg?Ala?Gln?Asn?Arg?Arg

105 110 115 120

ctg?ctg?ccc?gac?gcc?ccc?aac?gcc?cta?cac?atc?ctg?gac?ggg?ccc?gag 618

Leu?Leu?Pro?Asp?Ala?Pro?Asn?Ala?Leu?His?Ile?Leu?Asp?Gly?Pro?Glu

125 130 135

cag?cgc?tat?ttc?ctg?ggc?gtc?gtg?gat?ctc?gcc?aca?gtc?tac?ggg?ctc 666

Gln?Arg?Tyr?Phe?Leu?Gly?Val?Val?Asp?Leu?Ala?Thr?Val?Tyr?Gly?Leu

140 145 150

cgc?aag?cgg?ctg?gag?cac?ctg?tgg?aag?aca?ctg?cgc?tac?cca?ggc?cgg 714

Arg?Lys?Arg?Leu?Glu?His?Leu?Trp?Lys?Thr?Leu?Arg?Tyr?Pro?Gly?Arg

155 160 165

acc?ttc?tcc?act?gtc?agc?ccg?gct?cgc?tac?gcc?cgt?cgc?ctc?tgc?cag 762

Thr?Phe?Ser?Thr?Val?Ser?Pro?Ala?Arg?Tyr?Ala?Arg?Arg?Leu?Cys?Gln

170 175 180

tgg?gtg?gag?gcg?cac?acg?gag?tga?cgggcgcccg?gccccactct?ccggatctgg 816

Trp?Val?Glu?Ala?His?Thr?Glu

185 190

acgatgggct?cacgccagga?acgccggttc?ccccgggccc?gggcatctcg?cctgcgcttc 876

ctcctgatgg?tcgccagagg?gcagcatccc?ctaactaata?ccgtaaccgc?gcagtcccgt 936

ttgacggtgg?tgccgtgccc?agcatcgtgc?caaggactcc?cctactttag?ctcatttaat 996

cctcaaaaaa?tgctattatt?ctctttttac?agaccatgaa?atggaggctc?agggggtgaa 1056

gggactgatc?aagatcattc?agcaataaat?gctggaacca?ggactcaacc?atggcttttg 1116

gattccggag?cctgtattct?taaccaccag?ctcctgcagc?ttggtcctca?tgatctgggc 1176

aaggggggag?gctgaaggct?gcagccctct?tgtcatccag?atggggaaac?tgaggcccag 1236

agactttaag?gggcgtaagc?acgggtaagt?ggcaaggtcg?gccctgagta?cccaggcctc 1296

ccggccccct?gctcctggcc?tgatactcta?gggatgcagg?tgggagaagc?aggggtcctg 1356

ggggctgcct?ggagctctgg?gaggcattct?gaacggggtc?tactactgat?ctcaggtgag 1416

ctctgccctc?ctctgaaagt?cacttttctc?atcagttaaa?tgggggcaag?ggtccgtggt 1476

ccgaccaagg?tcttggcttc?acagacatca?ccaggagcct?gcatgcccct?gatcactcct 1536

tctccttcct?ccaggaaact?ccagcctggc?ctctgacccc?agttcaatcc?gaccatgccc 1596

aagcccaagc?gggcctttcc?tccagaactg?ctccggggcc?tggctgtgtg?actggagcaa 1656

ggtgctaaac?ctctctgtgc?ctcgctggtc?taatctgtaa?aatgaaggaa?tggaaacaga 1716

cctcattaac?tcattaaata?tttgttgagc?acctgcc 1753

<210>10

<211>191

<212>PRT

＜213〉people

<400>10

Met?Gln?Ser?Val?Phe?Tyr?Pro?Ala?Gly?Arg?Ile?Ser?Glu?Arg?Tyr?Asp

1 5 10 15

Ile?Lys?Gly?Cys?Glu?Val?Ser?Arg?Trp?Val?Asp?Pro?Ala?Pro?Glu?Gly

20 25 30

Ser?Pro?Leu?Val?Leu?Val?Leu?Lys?Asp?Leu?Asn?Phe?Gln?Gly?Lys?Thr

35 40 45

Ile?Asn?Leu?Gly?Pro?Gln?Arg?Ser?Trp?Phe?Leu?Arg?Gln?Met?Glu?Leu

50 55 60

Asp?Thr?Thr?Phe?Leu?Arg?Glu?Leu?Asn?Val?Leu?Asp?Tyr?Ser?Leu?Leu

65 70 75 80

Ile?Ala?Phe?Gln?Arg?Leu?His?Glu?Asp?Glu?Arg?Gly?Pro?Gly?Ser?Ser

85 90 95

Leu?Ile?Phe?Arg?Thr?Ala?Arg?Ser?Val?Gln?Gly?Ala?Gln?Ser?Pro?Glu

100 105 110

Glu?Ser?Arg?Ala?Gln?Asn?Arg?Arg?Leu?Leu?Pro?Asp?Ala?Pro?Asn?Ala

115 120 125

Leu?His?Ile?Leu?Asp?Gly?Pro?Glu?Gln?Arg?Tyr?Phe?Leu?Gly?Val?Val

130 135 140

Asp?Leu?Ala?Thr?Val?Tyr?Gly?Leu?Arg?Lys?Arg?Leu?Glu?His?Leu?Trp

145 150 155 160

Lys?Thr?Leu?Arg?Tyr?Pro?Gly?Arg?Thr?Phe?Ser?Thr?Val?Ser?Pro?Ala

165 170 175

Arg?Tyr?Ala?Arg?Arg?Leu?Cys?Gln?Trp?Val?Glu?Ala?His?Thr?Glu

180 185 190

<210>11

<211>2686

<212>DNA

＜213〉people

<220>

<221>CDS

<222>(78)..(1856)

<400>11

tcctcgatgc?caagaagagc?ctgaacctca?acatcttcct?gaagcaattt?aagtgctcca 60

acgaggaggt?cgctgct?atg?atc?cgg?gct?gga?gat?acc?acc?aag?ttt?gat 110

Met?Ile?Arg?Ala?Gly?Asp?Thr?Thr?Lys?Phe?Asp

1 5 10

gtg?gag?gtt?ctc?aaa?caa?ctc?ctt?aag?ctc?ctt?ccc?gag?aag?cac?gag 158

Val?Glu?Val?Leu?Lys?Gln?Leu?Leu?Lys?Leu?Leu?Pro?Glu?Lys?His?Glu

15 20 25

att?gaa?aac?ctg?cgg?gca?ttc?aca?gag?gag?cga?gcc?aag?ctg?gcc?agc 206

Ile?Glu?Asn?Leu?Arg?Ala?Phe?Thr?Glu?Glu?Arg?Ala?Lys?Leu?Ala?Ser

30 35 40

gcc?gac?cacttc?tac?ctc?ctc?ctg?ctg?gcc?att?ccc?tgc?tac?cag?ctg 254

Ala?Asp?His?Phe?Tyr?Leu?Leu?Leu?Leu?Ala?Ile?Pro?Cys?Tyr?Gln?Leu

45 50 55

cga?atc?gag?tgc?atg?ctg?ctg?tgt?gag?ggc?gcg?gcc?gcc?gtg?ctg?gac 302

Arg?Ile?Glu?Cys?Met?Leu?Leu?Cys?Glu?Gly?Ala?Ala?Ala?Val?Leu?Asp

60 65 70 75

atg?gtg?cgg?ccc?aag?gcc?cag?ctg?gtg?ctg?gct?gcc?tgc?gaa?agc?ctg 350

Met?Val?Arg?Pro?Lys?Ala?Gln?Leu?Val?Leu?Ala?Ala?Cys?Glu?Ser?Leu

80 85 90

ctc?acc?agc?cgc?cag?ctg?ccc?atc?ttc?tgc?cag?ctg?atc?ctg?aga?att 398

Leu?Thr?Ser?Arg?Gln?Leu?Pro?Ile?Phe?Cys?Gln?Leu?Ile?Leu?Arg?Ile

95 100 105

ggg?aac?ttc?ctc?aac?tac?ggc?agc?cac?acc?ggt?gac?gcc?gac?ggc?ttc 446

Gly?Asn?Phe?Leu?Asn?Tyr?Gly?Ser?His?Thr?Gly?Asp?Ala?Asp?Gly?Phe

110 115 120

aag?atc?agc?aca?ttg?ctg?aag?ctc?acg?gag?acc?aag?tcc?cag?cag?aac 494

Lys?Ile?Ser?Thr?Leu?Leu?Lys?Leu?Thr?Glu?Thr?Lys?Ser?Gln?Gln?Asn

125 130 135

cgc?gtg?acg?ctg?ctg?cac?cac?gtg?ctg?gag?gaa?gcg?gaa?aag?agc?cac 542

Arg?Val?Thr?Leu?Leu?His?His?Val?Leu?Glu?Glu?Ala?Glu?Lys?Ser?His

140 145 150 155

ccc?gac?ctc?ctg?cag?ctg?ccc?cgg?gac?ctg?gaa?cag?ccc?tcg?caa?gca 590

Pro?Asp?Leu?Leu?Gln?Leu?Pro?Arg?Asp?Leu?Glu?Gln?Pro?Ser?Gln?Ala

160 165 170

gca?ggg?atc?aac?ctg?gag?atc?atc?cgc?tca?gag?gcc?agc?tcc?aac?ctg 638

Ala?Gly?Ile?Asn?Leu?Glu?Ile?Ile?Arg?Ser?Glu?Ala?Ser?Ser?Asn?Leu

175 180 185

aag?aag?ctt?ctg?gag?acc?gag?cgg?aag?gtg?tct?gcc?tcc?gtg?gcc?gag 686

Lys?Lys?Leu?Leu?Glu?Thr?Glu?Arg?Lys?Val?Ser?Ala?Ser?Val?Ala?Glu

190 195 200

gtc?cag?gag?cag?tac?acc?gag?cgc?ctc?cag?gcc?agc?atc?tcg?gcc?ttc 734

Val?Gln?Glu?Gln?Tyr?Thr?Glu?Arg?Leu?Gln?Ala?Ser?Ile?Ser?Ala?Phe

205 210 215

cgg?gca?ctg?gac?gag?ctg?ttt?gag?gcc?atc?gag?cag?aag?caa?cgg?gag 782

Arg?Ala?Leu?Asp?Glu?Leu?Phe?Glu?Ala?Ile?Glu?Gln?Lys?Gln?Arg?Glu

220 225 230 235

ctg?gcc?gac?tac?ctg?tgt?gag?gac?gcc?cag?cag?ctg?tcc?ctg?gag?gac 830

Leu?Ala?Asp?Tyr?Leu?Cys?Glu?Asp?Ala?Gln?Gln?Leu?Ser?Leu?Glu?Asp

240 245 250

acg?ttc?agc?acc?atg?aag?gct?ttc?cgg?gac?ctt?ttc?ctc?cgc?gcc?ctg 878

Thr?Phe?Ser?Thr?Met?Lys?Ala?Phe?Arg?Asp?Leu?Phe?Leu?Arg?Ala?Leu

255 260 265

aag?gag?aac?aag?gac?cgg?aag?gag?cag?gcg?gcg?aag?gca?gag?agg?agg 926

Lys?Glu?Asn?Lys?Asp?Arg?Lys?Glu?Gln?Ala?Ala?Lys?Ala?Glu?Arg?Arg

270 275 280

aag?cag?cag?ctg?gcg?gag?gag?gag?gcg?cgg?cgg?cct?cgg?gga?gag?gac 974

Lys?Gln?Gln?Leu?Ala?Glu?Glu?Glu?Ala?Arg?Arg?Pro?Arg?Gly?Glu?Asp

285 290 295

ggg?aag?cct?gtc?agg?aag?ggg?ccc?ggg?aag?cag?gag?gag?gtg?tgt?gtc 1022

Gly?Lys?Pro?Val?Arg?Lys?Gly?Pro?Gly?Lys?Gln?Glu?Glu?Val?Cys?Val

300 305 310 315

atc?gat?gcc?ctg?ctg?gct?gac?atc?agg?aag?ggc?ttc?cag?ctg?cgg?aag 1070

Ile?Asp?Ala?Leu?Leu?Ala?Asp?Ile?Arg?Lys?Gly?Phe?Gln?Leu?Arg?Lys

320 325 330

aca?gcc?cgg?ggc?cgc?ggg?gac?acc?gac?ggg?ggc?agc?aag?gca?gcc?tcc 1118

Thr?Ala?Arg?Gly?Arg?Gly?Asp?Thr?Asp?Gly?Gly?Ser?Lys?Ala?Ala?Ser

335 340 345

atg?gat?ccc?cca?aga?gcc?aca?gag?cct?gtg?gcc?acc?agt?aac?cct?gca 1166

Met?Asp?Pro?Pro?Arg?Ala?Thr?Glu?Pro?Val?Ala?Thr?Ser?Asn?Pro?Ala

350 355 360

gga?gac?ccc?gtg?ggc?agc?acg?cgc?tgt?ccc?gcc?tct?gag?ccc?ggc?ctt 1214

Gly?Asp?Pro?Val?Gly?Ser?Thr?Arg?Cys?Pro?Ala?Ser?Glu?Pro?Gly?Leu

365 370 375

gat?gct?aca?aca?gcc?agc?gag?tcc?cgg?ggc?tgg?gac?ctt?gta?gac?gcc 1262

Asp?Ala?Thr?Thr?Ala?Ser?Glu?Ser?Arg?Gly?Trp?Asp?Leu?Val?Asp?Ala

380 385 390 395

gtg?acc?ccc?ggc?cct?cag?ccc?acc?ctg?gag?cag?ttg?gag?gag?ggt?ggt 1310

Val?Thr?Pro?Gly?Pro?Gln?Pro?Thr?Leu?Glu?Gln?Leu?Glu?Glu?Gly?Gly

400 405 410

ccg?cgg?ccc?ctg?gag?agg?cgt?tct?tcc?tgg?tat?gtg?gat?gcc?agc?gat 1358

Pro?Arg?Pro?Leu?Glu?Arg?Arg?Ser?Ser?Trp?Tyr?Val?Asp?Ala?Ser?Asp

415 420 425

gtc?cta?acc?act?gag?gat?ccc?cag?tgc?ccc?cag?ccc?ttg?gag?ggg?gcc 1406

Val?Leu?Thr?Thr?Glu?Asp?Pro?Gln?Cys?Pro?Gln?Pro?Leu?Glu?Gly?Ala

430 435 440

tgg?ccg?gtg?act?ctg?gga?gat?gct?cag?gcc?ctg?aag?ccc?ctc?aag?ttc 1454

Trp?Pro?Val?Thr?Leu?Gly?Asp?Ala?Gln?Ala?Leu?Lys?Pro?Leu?Lys?Phe

445 450 455

tcc?agc?aac?cag?ccc?cct?gca?gcc?gga?agt?tca?agg?caa?gat?gcc?aag 1502

Ser?Ser?Asn?Gln?Pro?Pro?Ala?Ala?Gly?Ser?Ser?Arg?Gln?Asp?Ala?Lys

460 465 470 475

gat?ccc?acg?tcc?ttg?ctg?ggc?gtc?ctc?cag?gcc?gag?gcc?gac?agc?aca 1550

Asp?Pro?Thr?Ser?Leu?Leu?Gly?Val?Leu?Gln?Ala?Glu?Ala?Asp?Ser?Thr

480 485 490

agt?gag?ggg?ctg?gag?gac?gct?gtc?cac?agc?cgt?ggt?gcc?aga?ccc?cct 1598

Ser?Glu?Gly?Leu?Glu?Asp?Ala?Val?His?Ser?Arg?Gly?Ala?Arg?Pro?Pro

495 500 505

gca?gca?ggc?cca?ggt?ggg?gat?gag?gac?gag?gac?gag?gag?gac?aca?gcc 1646

Ala?Ala?Gly?Pro?Gly?Gly?Asp?Glu?Asp?Glu?Asp?Glu?Glu?Asp?Thr?Ala

510 515 520

cca?gag?tcc?gca?ctg?gac?aca?tcc?ctg?gac?aag?tcc?ttc?tcc?gag?gat 1694

Pro?Glu?Ser?Ala?Leu?Asp?Thr?Ser?Leu?Asp?Lys?Ser?Phe?Ser?Glu?Asp

525 530 535

gcg?gtg?acc?gac?tcc?tcg?ggg?tcg?ggc?aca?ctc?ccc?agg?gcc?cgg?ggc 1742

Ala?Val?Thr?Asp?Ser?Ser?Gly?Ser?Gly?Thr?Leu?Pro?Arg?Ala?Arg?Gly

540 545 550 555

cgg?gcc?tca?aag?ggg?acc?ggg?aag?cga?agg?aag?aag?cgt?ccc?tcc?agg 1790

Arg?Ala?Ser?Lys?Gly?Thr?Gly?Lys?Arg?Arg?Lys?Lys?Arg?Pro?Ser?Arg

560 565 570

agc?cag?gaa?gag?gtt?ccc?cct?gat?tct?gat?gat?aat?aaa?aca?aag?aaa 1838

Ser?Gln?Glu?Glu?Val?Pro?Pro?Asp?Ser?Asp?Asp?Asn?Lys?Thr?Lys?Lys

575 580 585

ctg?tgt?gtg?atc?cag?taa?ggcctcaggc?ccaggcccaa?ggccaagtga 1886

Leu?Cys?Val?Ile?Gln

590

gagagcccag?gccacaggac?atgctgccat?tctgccaaga?gaggctcttc?tgggggccag 1946

gctgggactg?ggccccggaa?accaaaactc?cgtgccttac?ccagccgggg?ccctcctgga 2006

gccttcttgg?ggtgttgtgg?ctgggaaccc?gacaggcacc?agtgccctgc?caggcctggt 2066

gccctcctgg?accgcctgca?cgtgccagcc?tcccacctgc?ttcctaaagg?caaccctggc 2126

ccacacccgc?atgcgcccgg?tgcagcctgc?caagggccag?tcggggggtg?ctgcgtcctg 2186

ccagtgtcca?ccacagctct?gcctgccctt?cagcccagca?aggtttaatc?aaaatgcaat 2246

gctttgcaag?tctttactgc?ttggaggtgg?ctgagttggg?ggccctgggc?aggggtaagc 2306

tggcaggcag?tgccatggca?ggccagggtc?ccctcccatg?gggtctggcc?cccgttccag 2366

catgtccagc?ccctgaagtt?ggagttgggg?gcggtctgcc?tttgctgcca?ctgccaggcc 2426

tctgccctgc?agctgaaact?tggccatcac?atcaacagaa?aacccctccc?agtgccagct 2486

gcccagcgtg?ggcaggccct?ggggacaata?caggtccacc?tgaggggctg?cagggtgaca 2546

cccagcagcc?gctgccccct?cactgcccac?ccagcgaggg?cagcctaccc?gagcctgccc 2606

cctgccaggt?gtgtgccctg?aggctggcgg?ctggatgcgt?ggccaataaa?aagcagacct 2666

aaaaaaaaaa?aaaaaaaaaa 2686

<210>12

<211>592

<212>PRT

＜213〉people

<400>12

Met?Ile?Arg?Ala?Gly?Asp?Thr?Thr?Lys?Phe?Asp?Val?Glu?Val?Leu?Lys

1 5 10 15

Gln?Leu?Leu?Lys?Leu?Leu?Pro?Glu?Lys?His?Glu?Ile?Glu?Asn?Leu?Arg

20 25 30

Ala?Phe?Thr?Glu?Glu?Arg?Ala?Lys?Leu?Ala?Ser?Ala?Asp?His?Phe?Tyr

35 40 45

Leu?Leu?Leu?Leu?Ala?Ile?Pro?Cys?Tyr?Gln?Leu?Arg?Ile?Glu?Cys?Met

50 55 60

Leu?Leu?Cys?Glu?Gly?Ala?Ala?Ala?Val?Leu?Asp?Met?Val?Arg?Pro?Lys

65 70 75 80

Ala?Gln?Leu?Val?Leu?Ala?Ala?Cys?Glu?Ser?Leu?Leu?Thr?Ser?Arg?Gln

85 90 95

Leu?Pro?Ile?Phe?Cys?Gln?Leu?Ile?Leu?Arg?Ile?Gly?Asn?Phe?Leu?Asn

100 105 110

Tyr?Gly?Ser?His?Thr?Gly?Asp?Ala?Asp?Gly?Phe?Lys?Ile?Ser?Thr?Leu

115 120 125

Leu?Lys?Leu?Thr?Glu?Thr?Lys?Ser?Gln?Gln?Asn?Arg?Val?Thr?Leu?Leu

130 135 140

His?His?Val?Leu?Glu?Glu?Ala?Glu?Lys?Ser?His?Pro?Asp?Leu?Leu?Gln

145 150 155 160

Leu?Pro?Arg?Asp?Leu?Glu?Gln?Pro?Ser?Gln?Ala?Ala?Gly?Ile?Asn?Leu

165 170 175

Glu?Ile?Ile?Arg?Ser?Glu?Ala?Ser?Ser?Asn?Leu?Lys?Lys?Leu?Leu?Glu

180 185 190

Thr?Glu?Arg?Lys?Val?Ser?Ala?Ser?Val?Ala?Glu?Val?Gln?Glu?Gln?Tyr

195 200 205

Thr?Glu?Arg?Leu?Gln?Ala?Ser?Ile?Ser?Ala?Phe?Arg?Ala?Leu?Asp?Glu

210 215 220

Leu?Phe?Glu?Ala?Ile?Glu?Gln?Lys?Gln?Arg?Glu?Leu?Ala?Asp?Tyr?Leu

225 230 235 240

Cys?Glu?Asp?Ala?Gln?Gln?Leu?Ser?Leu?Glu?Asp?Thr?Phe?Ser?Thr?Met

245 250 255

Lys?Ala?Phe?Arg?Asp?Leu?Phe?Leu?Arg?Ala?Leu?Lys?Glu?Asn?Lys?Asp

260 265 270

Arg?Lys?Glu?Gln?Ala?Ala?Lys?Ala?Glu?Arg?Arg?Lys?Gln?Gln?Leu?Ala

275 280 285

Glu?Glu?Glu?Ala?Arg?Arg?Pro?Arg?Gly?Glu?Asp?Gly?Lys?Pro?Val?Arg

290 295 300

Lys?Gly?Pro?Gly?Lys?Gln?Glu?Glu?Val?Cys?Val?Ile?Asp?Ala?Leu?Leu

305 310 315 320

Ala?Asp?Ile?Arg?Lys?Gly?Phe?Gln?Leu?Arg?Lys?Thr?Ala?Arg?Gly?Arg

325 330 335

Gly?Asp?Thr?Asp?Gly?Gly?Ser?Lys?Ala?Ala?Ser?Met?Asp?Pro?Pro?Arg

340 345 350

Ala?Thr?Glu?Pro?Val?Ala?Thr?Ser?Asn?Pro?Ala?Gly?Asp?Pro?Val?Gly

355 360 365

Ser?Thr?Arg?Cys?Pro?Ala?Ser?Glu?Pro?Gly?Leu?Asp?Ala?Thr?Thr?Ala

370 375 380

Ser?Glu?Ser?Arg?Gly?Trp?Asp?Leu?Val?Asp?Ala?Val?Thr?Pro?Gly?Pro

385 390 395 400

Gln?Pro?Thr?Leu?Glu?Gln?Leu?Glu?Glu?Gly?Gly?Pro?Arg?Pro?Leu?Glu

405 410 415

Arg?Arg?Ser?Ser?Trp?Tyr?Val?Asp?Ala?Ser?Asp?Val?Leu?Thr?Thr?Glu

420 425 430

Asp?Pro?Gln?Cys?Pro?Gln?Pro?Leu?Glu?Gly?Ala?Trp?Pro?Val?Thr?Leu

435 440 445

Gly?Asp?Ala?Gln?Ala?Leu?Lys?Pro?Leu?Lys?Phe?Ser?Ser?Asn?Gln?Pro

450 455 460

Pro?Ala?Ala?Gly?Ser?Ser?Arg?Gln?Asp?Ala?Lys?Asp?Pro?Thr?Ser?Leu

465 470 475 480

Leu?Gly?Val?Leu?Gln?Ala?Glu?Ala?Asp?Ser?Thr?Ser?Glu?Gly?Leu?Glu

485 490 495

Asp?Ala?Val?His?Ser?Arg?Gly?Ala?Arg?Pro?Pro?Ala?Ala?Gly?Pro?Gly

500 505 510

Gly?Asp?Glu?Asp?Glu?Asp?Glu?Glu?Asp?Thr?Ala?Pro?Glu?Ser?Ala?Leu

515 520 525

Asp?Thr?Ser?Leu?Asp?Lys?Ser?Phe?Ser?Glu?Asp?Ala?Val?Thr?Asp?Ser

530 535 540

Ser?Gly?Ser?Gly?Thr?Leu?Pro?Arg?Ala?Arg?Gly?Arg?Ala?Ser?Lys?Gly

545 550 555 560

Thr?Gly?Lys?Arg?Arg?Lys?Lys?Arg?Pro?Ser?Arg?Ser?Gln?Glu?Glu?Val

565 570 575

Pro?Pro?Asp?Ser?Asp?Asp?Asn?Lys?Thr?Lys?Lys?Leu?Cys?Val?Ile?Gln

580 585 590

<210>13

<211>2190

<212>DNA

＜213〉people

<220>

<221>CDS

<222>(148)..(702)

<400>13

cggacgcgtg?gggagaggct?gtttaccaga?acagcataac?aagggcaggt?ctgactgcaa 60

ggctgggact?gggaggcaga?gccgccgcca?agggggcctc?ggttaaacac?tggtcgttca 120

atcacctgca?agacgaagga?ggcaagg?atg?ctg?ttg?gcc?tgg?gta?caa?gca?ttc 174

Met?Leu?Leu?Ala?Trp?Val?Gln?Ala?Phe

1 5

ctc?gtc?agc?aac?atg?ctc?cta?gca?gaa?gcc?tat?gga?tct?gga?gag?acc 222

Leu?Val?Ser?Asn?Met?Leu?Leu?Ala?Glu?Ala?Tyr?Gly?Ser?Gly?Glu?Thr

10 15 20 25

acc?tcc?cag?gcc?ctg?cca?gcc?ttc?acg?aca?gaa?atc?cag?gaa?gcg?tct 270

Thr?Ser?Gln?Ala?Leu?Pro?Ala?Phe?Thr?Thr?Glu?Ile?Gln?Glu?Ala?Ser

30 35 40

gaa?ggg?cca?ggt?gca?gat?gag?gtg?cag?gtg?ttc?gct?cct?gcc?aac?gcc 318

Glu?Gly?Pro?Gly?Ala?Asp?Glu?Val?Gln?ValPhe?Ala?Pro?Ala?Asn?Ala

45 50 55

ctg?ccc?gct?cgg?agt?gag?gcg?gca?gct?gtg?cag?cca?gtg?att?ggg?atc 366

Leu?Pro?Ala?Arg?Ser?Glu?Ala?Ala?Ala?Val?Gln?Pro?Val?Ile?Gly?Ile

60 65 70

agc?cag?cgg?gtg?cgg?atg?aac?tcc?aag?gag?aaa?aag?gac?ctg?gga?act 414

Ser?Gln?Arg?Val?Arg?Met?Asn?Ser?Lys?Glu?Lys?Lys?Asp?Leu?Gly?Thr

75 80 85

ctg?ggc?tac?gtg?ctg?ggc?att?acc?atg?atg?gtg?atc?atc?att?gcc?atc 462

Leu?Gly?Tyr?Val?Leu?Gly?Ile?Thr?Met?Met?Val?Ile?Ile?Ile?Ala?Ile

90 95 100 105

gga?gct?ggc?atc?atc?ttg?ggc?tac?tcc?tac?aag?agg?ggg?aag?gat?ttg 510

Gly?Ala?Gly?Ile?Ile?Leu?Gly?Tyr?Ser?Tyr?Lys?Arg?Gly?Lys?Asp?Leu

110 115 120

aaa?gaa?cag?cat?gat?cag?aaa?gta?tgt?gag?agg?gag?atg?cag?cga?atc 558

Lys?Glu?Gln?His?Asp?Gln?Lys?Val?Cys?Glu?Arg?Glu?Met?Gln?Arg?Ile

125 130 135

act?ctg?ccc?ttg?tct?gcc?ttc?acc?aac?ccc?acc?tgt?gag?att?gtg?gat 606

Thr?Leu?Pro?Leu?Ser?Ala?Phe?Thr?Asn?Pro?Thr?Cys?Glu?Ile?Val?Asp

140 145 150

gag?aag?act?gtc?gtg?gtc?cac?acc?agc?cag?act?cca?gtt?gac?cct?cag 654

Glu?Lys?Thr?Val?Val?Val?His?Thr?Ser?Gln?Thr?Pro?Val?Asp?Pro?Gln

155 160 165

gag?ggc?acc?acc?ccc?ctt?atg?ggc?cag?gcc?ggg?act?cct?ggg?gcc?tga 702

Glu?Gly?Thr?Thr?Pro?Leu?Met?Gly?Gln?Ala?Gly?Thr?Pro?Gly?Ala

170 175 180

gcccccccag?tgggcaggag?cccatgcaga?cactggtgca?ggacagccca?ccctcctaca 762

gctaggagga?actaccactt?tgtgttctgg?ttaaaaccct?accactcccc?cgcttttttg 822

gcgaatccta?gtaagagtga?cagaagcagg?tggccctgtg?ggctgagggt?aaggctgggt 882

agggtcctaa?cagtgctcct?tgtccatccc?ttggagcaga?ttttgtctgt?ggatggagac 942

agtggcagct?cccacagtga?tgctgctgct?aagggcttcc?aaacattgcc?tgcacccctg 1002

gaactgaacc?agggatagac?ggggagctcc?cccaggctcc?tctgtgcttt?actaagatgg 1062

cctcagtctc?cactgtgggc?ttgagtggca?tacactgtta?ttcatggtta?aggtaaagca 1122

ggtcaaggga?tggcattgaa?aaaatatatt?tagtttttaa?aatatttggg?atggaactcc 1182

ctactgacct?ctgagaactg?gaaacgagtt?tgtacagaag?tcagaacttt?gggttgggaa 1242

tgagatctag?gttgtggctg?ctggtatgct?tcagcttgct?ggcaatgatg?tgccttgaca 1302

accgtgggcc?aggcctgggc?ccagggactc?ttcctgtttc?ataaggaaag?gaagaattgc 1362

actgagcatt?ccacttagga?agaggataga?gaaggatctgctccgccttt?ggccacagga 1422

gcagaggcag?acctgggatg?ccccagtttc?tcttcaggga?tggatagtga?cctgtcttca 1482

ttttgcacag?gtaagagagt?agttagctaa?cctatgggaa?ttatactgtg?gggccttgtg 1542

agctgcttct?aagaggctaa?cctggaaact?aagctcagag?gcaaggtaat?aaagcacttc 1602

agggcttgct?ccccaagtgg?gcctgattta?gcaggtggtc?ctgcgggcgt?ccaggtcagc 1662

accttcctgt?agggcactgg?ggctagggtc?acagccccta?actcataaag?caatcaaaga 1722

accattagaa?agggctcatt?aagccttttg?gacacaggac?cccagagagg?aaaaagtgac 1782

ttgcccaagg?tcgtaagcaa?gctactggca?tggcaagagc?ccagcttcct?gacggagcgc 1842

aacatttctc?cactgcactg?tgctagcagc?tcagcagggc?ctctaacctg?tgatgtcaca 1902

ctcaagaggc?cttggcagct?cctagccata?gagcttcctt?tccagaaccc?ttccactgcc 1962

caatgtggag?acaggggtta?gtggggcttt?ctatggagcc?atctgctttg?gggacctaga 2022

cctcaggtgg?tctcttggtg?ttagtgatgc?tggagaagag?aatattactg?gtttctactt 2082

ttctataaag?gcatttctct?atatacatgt?tttatatacc?tcattctgac?acctgcatat 2142

agtgtgggaa?attgctctgc?atttgactta?attaaaaaaa?aaaaaaaa 2190

<210>14

<211>184

<212>PRT

＜213〉people

<400>14

Met?Leu?Leu?Ala?Trp?Val?Gln?Ala?Phe?Leu?Val?Ser?Asn?Met?Leu?Leu

1 5 10 15

Ala?Glu?Ala?Tyr?Gly?Ser?Gly?Glu?Thr?Thr?Ser?Gln?Ala?Leu?Pro?Ala

20 25 30

Phe?Thr?Thr?Glu?Ile?Gln?Glu?Ala?Ser?Glu?Gly?Pro?Gly?Ala?Asp?Glu

35 40 45

Val?Gln?Val?Phe?Ala?Pro?Ala?Asn?Ala?Leu?Pro?Ala?Arg?Ser?Glu?Ala

50 55 60

Ala?Ala?Val?Gln?Pro?Val?Ile?Gly?Ile?Ser?Gln?Arg?Val?Arg?Met?Asn

65 70 75 80

Ser?Lys?Glu?Lys?Lys?Asp?Leu?Gly?Thr?Leu?Gly?Tyr?Val?Leu?Gly?Ile

85 90 95

Thr?Met?Met?Val?Ile?Ile?Ile?Ala?Ile?Gly?Ala?Gly?Ile?Ile?Leu?Gly

100 105 110

Tyr?Ser?Tyr?Lys?Arg?Gly?Lys?Asp?Leu?Lys?Glu?Gln?His?Asp?Gln?Lys

115 120 125

Val?Cys?Glu?Arg?Glu?Met?Gln?Arg?Ile?Thr?Leu?Pro?Leu?Ser?Ala?Phe

130 135 140

Thr?Asn?Pro?Thr?Cys?Glu?Ile?Val?Asp?Glu?Lys?Thr?Val?Val?Val?His

145 150 155 160

Thr?Ser?Gln?Thr?Pro?Val?Asp?Pro?Gln?Glu?Gly?Thr?Thr?Pro?Leu?Met

165 170 175

Gly?Gln?Ala?Gly?Thr?Pro?Gly?Ala

180

<210>15

<211>1817

<212>DNA

＜213〉people

<220>

<221>CDS

<222>(439)..(897)

<400>15

agcataacct?gtttgtgaaa?ctgcaaaaag?gttgacatgg?ctgaagcaga?agatgggttg 60

gtactgaatg?gttgggagag?atatgtagcg?gctgaccctg?aaggacgttg?cagcccatgc 120

tatggagacc?tgattttatt?ttgtaagagg?tgggcaaccc?ctgaagcatt?ttagtgatgc 180

gaacagatgt?gtctgttacc?aagcaaagcc?agttaaagaa?ggtcacagag?gaggaatgat 240

cacaaagagt?ctgagaagcc?aactggggcc?aaacaagttt?tagaacctct?actggctgat 300

gccaacatca?tcagagatga?actgttctta?ccagaaactt?ctctgctcct?gacaccccac 360

ctctccccag?aagagaagaa?ggagagtggc?cacgttgatt?cagccaagca?cctccaggag 420

gtcccctctg?gatgtccc?atg?agg?ctg?ccc?ctc?agc?cac?agc?cca?gag?cac 471

Met?Arg?Leu?Pro?Leu?Ser?His?Ser?Pro?Glu?His

1 5 10

gtg?gag?atg?gct?ttg?ctc?agc?aac?atc?cta?gcg?gcc?tat?tcc?ttt?gtc 519

Val?Glu?Met?Ala?Leu?Leu?Ser?Asn?Ile?Leu?Ala?Ala?Tyr?Ser?Phe?Val

15 20 25

tca?gaa?aat?cct?gag?cga?gca?gct?ctg?tac?ttt?gtt?tct?ggc?gtg?tgc 567

Ser?Glu?Asn?Pro?Glu?Arg?Ala?Ala?Leu?Tyr?Phe?Val?Ser?Gly?Val?Cys

30 35 40

atc?ggg?ctg?gtg?ctg?acc?ctg?gct?gct?ctg?gtg?ata?agg?atc?tct?tgc 615

Ile?Gly?Leu?Val?Leu?Thr?Leu?Ala?Ala?Leu?Val?Ile?Arg?Ile?Ser?Cys

45 50 55

cac?aca?gac?tgc?agg?cgg?cgt?ccc?ggg?aag?aag?ttc?ctg?cag?gac?aga 663

His?Thr?Asp?Cys?Arg?Arg?Arg?Pro?Gly?Lys?Lys?Phe?Leu?Gln?Asp?Arg

60 65 70 75

gag?agc?agc?agc?gac?agc?agc?gac?agc?gag?gat?ggc?agt?gag?gac?acc 711

Glu?Ser?Ser?Ser?Asp?Ser?Ser?Asp?Ser?Glu?Asp?Gly?Ser?Glu?Asp?Thr

80 85 90

gtg?tcc?gat?ctc?tcc?gtg?cgg?aga?cac?cgc?cgc?ttc?gag?agg?act?ttg 759

Val?Ser?Asp?Leu?Ser?Val?Arg?Arg?His?Arg?Arg?Phe?Glu?Arg?Thr?Leu

95 100 105

aac?aag?aat?gtg?ttc?acc?tct?gcg?gag?gag?ctg?gag?cgc?gcc?cag?cgg 807

Asn?Lys?Asn?Val?Phe?Thr?Ser?Ala?Glu?Glu?Leu?Glu?Arg?Ala?Gln?Arg

110 115 120

ctg?gag?gag?cgc?gag?cgc?atc?atc?agg?gag?atc?tgg?atg?aat?ggc?cag 855

Leu?Glu?Glu?Arg?Glu?Arg?Ile?Ile?Arg?Glu?Ile?Trp?Met?Asn?Gly?Gln

125 130 135

cct?gag?gtg?ccc?ggg?acc?agg?agc?ctg?aat?cgc?tac?tat?tag 897

Pro?Glu?Val?Pro?Gly?Thr?Arg?Ser?Leu?Asn?Arg?Tyr?Tyr

140 145 150

ggagcagcag?gaccccggaa?accactggag?gccgcctgga?aaggagagcg?tctgcaggga 957

cagtgggcac?aaggaactga?acccagctct?gctaatattg?tgatttcaga?gaaaaagcag 1017

gacatgcccc?ttttctagcc?aggaggattg?ctcctttttg?gccaaatgta?tggagaagta 1077

gaaaaatcaa?agcagttcat?caccctttcc?aggtctcgga?atggtgctga?aaaatcctct 1137

ccaacactgt?ggatggagat?cggagaaacg?cgactgtggt?ttctcttgat?ttctgaggat 1197

ctcagaagtt?tcagcagact?tccttgctct?gtgttattcc?tttgcaaaag?gaaagatcat 1257

tatagcatga?gggctgggaa?tagcagggtg?aacttaaccc?aataaatgca?atttcctaaa 1317

tgacttcatg?ctatggaggt?gattctgatt?caaatagcat?gatccatgct?tattatttaa 1377

ggctgatttt?taaaatcttg?tgttgcaagg?gcaacctcgt?ccattttaac?tggcacctca 1437

gggattaaaa?tcctactttt?tagtgtagtt?cccacctcat?tcatgaaaat?gaatagaact 1497

attgtattat?gggagatgtg?tcagtgaact?agaaaatgtc?aataacctca?aaggatgaag 1557

ggtttttatt?ttaaatagtt?tataaaatat?atattgacat?gcatatttga?aaatgctgca 1617

taaaaataaa?agtggtgtgt?tttccccctc?tttggagaca?agaaaatttt?attatgactc 1677

tagataattg?tgattttaaa?cactttgttt?tttttttttt?ttttttaatt?ggatttcaaa 1737

gaaaagaatg?gaaatgagag?gtaaggatta?aagccaaagt?taggatggga?attaaaaaaa 1797

taaacattac?ataaaatctt 1817

<210>16

<211>152

<212>PRT

＜213〉people

<400>16

Met?Arg?Leu?Pro?Leu?Ser?His?Ser?Pro?Glu?His?Val?Glu?Met?Ala?Leu

1 5 10 15

Leu?Ser?Asn?Ile?Leu?Ala?Ala?Tyr?Ser?Phe?Val?Ser?Glu?Asn?Pro?Glu

20 25 30

Arg?Ala?Ala?Leu?Tyr?Phe?Val?Ser?Gly?Val?Cys?Ile?Gly?Leu?Val?Leu

35 40 45

Thr?Leu?Ala?Ala?Leu?Val?Ile?Arg?Ile?Ser?Cys?His?Thr?Asp?Cys?Arg

50 55 60

Arg?Arg?Pro?Gly?Lys?Lys?Phe?Leu?Gln?Asp?Arg?Glu?Ser?Ser?Ser?Asp

65 70 75 80

Ser?Ser?Asp?Ser?Glu?Asp?Gly?Ser?Glu?Asp?Thr?Val?Ser?Asp?Leu?Ser

85 90 95

Val?Arg?Arg?His?Arg?Arg?Phe?Glu?Arg?Thr?Leu?Asn?Lys?Asn?Val?Phe

100 105 110

Thr?Ser?Ala?Glu?Glu?Leu?Glu?Arg?Ala?Gln?Arg?Leu?Glu?Glu?Arg?Glu

115 120 125

Arg?Ile?Ile?Arg?Glu?Ile?Trp?Met?Asn?Gly?Gln?Pro?Glu?Val?Pro?Gly

130 135 140

Thr?Arg?Ser?Leu?Asn?Arg?Tyr?Tyr

145 150

<210>17

<211>1868

<212>DNA

＜213〉people

<220>

<221>CDS

<222>(403)..(870)

<400>17

agtttcggaa?ccccagccag?ctcacctctg?cgccgctgaa?cccgatccga?gcctccggca 60

aaggtttttc?cctcctcccc?cggccgaggg?cttctgccgc?ccgggcaccc?ccgccccgcg 120

gcgccccaca?ttcccccagc?ccggggccct?tggcgcgtgc?gctccgtgcg?gctgtgctcc 180

gcgggacttt?gtttgtttcc?tcctcgtccc?tctttgttgg?gctgaacacc?agcctcgtca 240

aagcccccca?ctccggaggg?agttcggctt?ctccagcagg?gcggctgcag?cgcgctgccc 300

cgaccccgcc?tgcggcccct?cacgccgcta?gtgctcccac?cccgccctcc?tggcaccccg 360

cctgcgtccg?ttcgcccgag?gaagccaacc?gcgacttcat?tg?atg?cac?cca?ttc 414

Met?His?Pro?Phe

1

cag?tgg?tgt?aac?ggg?tgt?ttc?tgt?ggc?ctg?gga?ctg?gtt?agc?acc?aac 462

Gln?Trp?Cys?Asn?Gly?Cys?Phe?Cys?Gly?Leu?Gly?Leu?Val?Ser?Thr?Asn

5 10 15 20

aag?tcc?tgc?tcg?atg?cca?ccc?atc?agt?ttc?cag?gac?ctt?ccg?ctc?aac 510

Lys?Ser?Cys?Ser?Met?Pro?Pro?Ile?Ser?Phe?Gln?Asp?Leu?Pro?Leu?Asn

25 30 35

atc?tat?atg?gtc?atc?ttc?ggc?aca?ggc?atc?ttt?gtc?ttc?atg?ctc?agc 558

Ile?Tyr?Met?Val?Ile?Phe?Gly?Thr?Gly?Ile?Phe?Val?Phe?Met?Leu?Ser

40 45 50

ctt?atc?ttc?tgc?tgc?tat?ttt?atc?agc?aaa?ctg?cgg?aac?cag?gca?cag 606

Leu?Ile?Phe?Cys?Cys?Tyr?Phe?Ile?Ser?Lys?Leu?Arg?Asn?Gln?Ala?Gln

55 60 65

agt?gag?cga?tac?gga?tat?aag?gag?gtg?gtg?ctt?aaa?ggt?gat?gcc?aag 654

Ser?Glu?Arg?Tyr?Gly?Tyr?Lys?Glu?Val?Val?Leu?Lys?Gly?Asp?Ala?Lys

70 75 80

aag?tta?caa?tta?tat?ggg?cag?acc?tgc?gca?gtc?tgt?ctg?gaa?gac?ttc 702

Lys?Leu?Gln?Leu?Tyr?Gly?Gln?Thr?Cys?Ala?Val?Cys?Leu?Glu?Asp?Phe

85 90 95 100

aag?ggg?aag?gat?gag?tta?ggc?gtg?ctc?ccg?tgc?caa?cac?gcc?ttt?cac 750

Lys?Gly?Lys?Asp?Glu?Leu?Gly?Val?Leu?Pro?Cys?Gln?His?Ala?Phe?His

105 110 115

cgc?aag?tgt?ctg?gtg?aaa?tgg?ctg?gaa?gtt?cgc?tgt?gtc?tgc?ccc?atg 798

Arg?Lys?Cys?Leu?Val?Lys?Trp?Leu?Glu?Val?Arg?Cys?Val?Cys?Pro?Met

120 125 130

tgt?aac?aag?ccc?att?gct?agt?ccc?tca?gag?gcc?acg?cag?aac?att?ggg 846

Cys?Asn?Lys?Pro?Ile?Ala?Ser?Pro?Ser?Glu?Ala?Thr?Gln?Asn?Ile?Gly

135 140 145

att?cta?ttg?gat?gag?ctg?gtg?tga?gtgctgccgc?tacaccgaga?cctggagaag 900

Ile?Leu?Leu?Asp?Glu?Leu?Val

150 155

acctcttgcc?tcatggatgt?ctggtccctc?tgcacagctc?caaccaacag?gactgtaggg 960

tgatgacgat?cactttccca?gtgatgagaa?gggtggtcta?ggactgggct?tctaccctca 1020

gtgcaagacc?agtgccagat?gtgcccccac?ttcctgcctc?ctgaagcctt?cttccctgct 1080

actccatgct?ggtggcctca?cccatcaaga?ccactgtctc?ctggtactgg?actatctacc 1140

tgccttgtcc?ctgttctggg?ggaaggtgtc?caccccgatc?aagaacatgg?agaaagtcct 1200

ctttcaaggc?tcccattagg?aggatgagct?gccttgaccc?agaagggatg?agacgggctc 1260

ttacctctct?acaaccttcc?ctccccttcc?cactccttcc?ggagtaaggt?tagaagggaa 1320

ggaaggaaag?atcaaggaac?caagcgcctc?cacgggaggc?gagggaggct?ctgtatgaaa 1380

cagaagagca?gggacataaa?ggaaaatgtc?agtgtttaca?tgggacctat?ggaaacaaag 1440

gctggcgggc?gccagctgac?tccagagtaa?gagagggccc?ttcccctgcc?aggacccacg 1500

gtgctatcca?ttcagtctct?tcctcagtta?atctcggagc?ttcctattcc?atgttgaggt 1560

ttgtgggccc?ctctagagga?gggctagttc?tatacttaaa?ttgattccca?ggggcctttt 1620

tttttttttt?tttttttttt?ttgatcaaaa?ggggtgtggg?gatgggggtg?tctacggtta 1680

agcaacagat?acctccttcc?ctttgtaaat?agtattttta?tacttcatcc?tcgcctctca 1740

ggctttagat?acgaaatctc?cagaatggaa?gggggtgggg?attttctgtt?cctccctgga 1800

gtgggtgagg?gtgggagaaa?gttacatatt?taaagaaaaa?taaatttaat?aacaagtttc 1860

tctaacct 1868

<210>18

<211>155

<212>PRT

＜213〉people

<400>18

Met?His?Pro?Phe?Gln?Trp?Cys?Asn?Gly?Cys?Phe?Cys?Gly?Leu?Gly?Leu

1 5 10 15

Val?Ser?Thr?Asn?Lys?Ser?Cys?Ser?Met?Pro?Pro?Ile?Ser?Phe?Gln?Asp

20 25 30

Leu?Pro?Leu?Asn?Ile?Tyr?Met?Val?Ile?Phe?Gly?Thr?Gly?Ile?Phe?Val

35 40 45

Phe?Met?Leu?Ser?Leu?Ile?Phe?Cys?Cys?Tyr?Phe?Ile?Ser?Lys?Leu?Arg

50 55 60

Asn?Gln?Ala?Gln?Ser?Glu?Arg?Tyr?Gly?Tyr?Lys?Glu?Val?Val?Leu?Lys

65 70 75 80

Gly?Asp?Ala?Lys?Lys?Leu?Gln?Leu?Tyr?Gly?Gln?Thr?Cys?Ala?Val?Cys

85 90 95

Leu?Glu?Asp?Phe?Lys?Gly?Lys?Asp?Glu?Leu?Gly?Val?Leu?Pro?Cys?Gln

100 105 110

His?Ala?Phe?His?Arg?Lys?Cys?Leu?Val?Lys?Trp?Leu?Glu?Val?Arg?Cys

115 120 125

Val?Cys?Pro?Met?Cys?Asn?Lys?Pro?Ile?Ala?Ser?Pro?Ser?Glu?Ala?Thr

130 135 140

Gln?Asn?Ile?Gly?Ile?Leu?Leu?Asp?Glu?Leu?Val

145 150 155

<210>19

<211>2651

<212>DNA

＜213〉people

<220>

<221>CDS

<222>(196)..(1599)

<400>19

ggcacgaggg?gacgcgataa?atatgcagag?cggaggcttc?gcgcagcaga?gcccgcgcgc 60

cgcccgctcc?gggtgctgaa?tccaggcgtg?gggacacgag?ccaggcgccg?ccgccggagc 120

cagcggagcc?ggggccagag?ccggagcgcg?tccgcgtcca?cgcagccgcc?ggccggccag 180

cacccagggc?cctgc?atg?cca?ggt?cgt?tgg?agg?tgg?cag?cga?gac?atg?cac 231

Met?Pro?Gly?Arg?Trp?Arg?Trp?Gln?Arg?Asp?Met?His

1 5 10

ccg?gcc?cgg?aag?ctc?ctc?agc?ctc?ctc?ttc?ctc?atc?ctg?atg?ggc?act 279

Pro?Ala?Arg?Lys?Leu?Leu?Ser?Leu?Leu?Phe?Leu?Ile?Leu?Met?Gly?Thr

15 20 25

gaa?ctc?act?caa?gtg?ctg?ccc?acc?aac?cct?gag?gag?agc?tgg?cag?gtg 327

Glu?Leu?Thr?Gln?Val?Leu?Pro?Thr?Asn?Pro?Glu?Glu?Ser?Trp?Gln?Val

30 35 40

tac?agc?tct?gcc?cag?gac?agc?gag?ggc?agg?tgt?atc?tgc?aca?gtg?gtc 375

Tyr?Ser?Ser?Ala?Gln?Asp?Ser?Glu?Gly?Arg?Cys?Ile?Cys?Thr?Val?Val

45 50 55 60

gcc?cca?cag?cag?acc?atg?tgt?tca?cgg?gat?gcc?cgc?aca?aaa?cag?ctg 423

Ala?Pro?Gln?Gln?Thr?Met?Cys?Ser?Arg?Asp?Ala?Arg?Thr?Lys?Gln?Leu

65 70 75

agg?cag?cta?ctg?gag?aag?gtg?cag?aac?atg?tct?caa?tcc?ata?gag?gtc 471

Arg?Gln?Leu?Leu?Glu?Lys?Val?Gln?Asn?Met?Ser?Gln?Ser?Ile?Glu?Val

80 85 90

ttg?gac?agg?cgg?acc?cag?aga?gac?ttg?cag?tac?gtg?gag?aag?atg?gag 519

Leu?Asp?Arg?Arg?Thr?Gln?Arg?Asp?Leu?Gln?Tyr?Val?Glu?Lys?Met?Glu

95 100 105

aac?caa?atg?aaa?gga?ctg?gag?tcc?aag?ttc?aaa?cag?gtg?gag?gag?agt 567

Asn?Gln?Met?Lys?Gly?Leu?Glu?Ser?Lys?Phe?Lys?Gln?Val?Glu?Glu?Ser

110 115 120

cat?aag?caa?cac?ctg?gcc?agg?cag?ttt?aag?gcg?ata?aaa?gcg?aaa?atg 615

His?Lys?Gln?His?Leu?Ala?Arg?Gln?Phe?Lys?Ala?Ile?Lys?Ala?Lys?Met

125 130 135 140

gat?gaa?ctt?agg?cct?ttg?ata?cct?gtg?ttg?gaa?gag?tac?aag?gcc?gat 663

Asp?Glu?Leu?Arg?Pro?Leu?Ile?Pro?Val?Leu?Glu?Glu?Tyr?Lys?Ala?Asp

145 150 155

gcc?aaa?ttg?gta?ttg?cag?ttt?aaa?gag?gag?gtc?cag?aat?ctg?acg?tca 711

Ala?Lys?Leu?Val?Leu?Gln?Phe?Lys?Glu?Glu?Val?Gln?Asn?Leu?Thr?Ser

160 165 170

gtg?ctt?aac?gag?ctg?caa?gag?gaa?att?ggc?gcc?tat?gac?tac?gat?gaa 759

Val?Leu?Asn?Glu?Leu?Gln?Glu?Glu?Ile?Gly?Ala?Tyr?Asp?Tyr?Asp?Glu

175 180 185

ctt?cag?agc?aga?gtg?tcc?aat?ctt?gaa?gaa?agg?ctc?cgt?gca?tgc?atg 807

Leu?Gln?Ser?Arg?Val?Ser?Asn?Leu?Glu?Glu?Arg?Leu?Arg?Ala?Cys?Met

190 195 200

caa?aaa?cta?gct?tgc?ggg?aag?ttg?acg?ggc?atc?agt?gac?ccc?gtg?act 855

Gln?Lys?Leu?Ala?Cys?Gly?Lys?Leu?Thr?Gly?Ile?Ser?Asp?Pro?Val?Thr

205 210 215 220

gtc?aag?acc?tcc?ggc?tcg?agg?ttc?gga?tcc?tgg?atg?aca?gac?cct?ctc 903

Val?Lys?Thr?Ser?Gly?Ser?Arg?Phe?Gly?Ser?Trp?Met?Thr?Asp?Pro?Leu

225 230 235

gcc?cct?gaa?ggc?gat?aac?cgg?gtg?tgg?tac?atg?gac?ggc?tat?cac?aac 951

Ala?Pro?Glu?Gly?Asp?Asn?Arg?Val?Trp?Tyr?Met?Asp?Gly?Tyr?His?Asn

240 245 250

aac?cgc?ttc?gta?cgt?gag?tac?aag?tcc?atg?gtt?gac?ttc?atg?aac?acg 999

Asn?Arg?Phe?Val?Arg?Glu?Tyr?Lys?Ser?Met?Val?Asp?Phe?Met?Asn?Thr

255 260 265

gac?aat?ttc?acc?tcc?cac?cgt?ctc?ccc?cac?ccc?tgg?tcg?ggc?acg?ggg 1047

Asp?Asn?Phe?Thr?Ser?His?Arg?Leu?Pro?His?Pro?Trp?Ser?Gly?Thr?Gly

270 275 280

cag?gtg?gtc?tac?aac?ggt?tct?atc?tacttc?aac?aag?ttc?cag?agc?cac 1095

Gln?Val?Val?Tyr?Asn?Gly?Ser?Ile?Tyr?Phe?Asn?Lys?Phe?Gln?Ser?His

285 290 295 300

atc?atc?atc?agg?ttt?gac?ctg?aag?aca?gag?acc?atc?ctc?aag?acc?cgc 1143

Ile?Ile?Ile?Arg?Phe?Asp?Leu?Lys?Thr?Glu?Thr?Ile?Leu?Lys?Thr?Arg

305 310 315

agc?ctg?gac?tat?gcc?ggt?tac?aac?aac?atg?tac?cac?tac?gcc?tgg?ggt 1191

Ser?Leu?Asp?Tyr?Ala?Gly?Tyr?Asn?Asn?Met?Tyr?His?Tyr?Ala?Trp?Gly

320 325 330

ggc?cac?tcg?gac?atc?gac?ctc?atg?gtg?gac?gag?agc?ggg?ctg?tgg?gcc 1239

Gly?His?Ser?Asp?Ile?Asp?Leu?Met?Val?Asp?Glu?Ser?Gly?Leu?Trp?Ala

335 340 345

gtg?tac?gcc?acc?aac?cag?aac?gct?ggc?aac?atc?gtg?gtc?agt?agg?ctg 1287

Val?Tyr?Ala?Thr?Asn?Gln?Asn?Ala?Gly?Asn?Ile?Val?Val?Ser?Arg?Leu

350 355 360

gac?ccc?gtg?tcc?ctg?cag?acc?ctg?cag?acc?tgg?aac?acg?agc?tac?ccc 1335

Asp?Pro?Val?Ser?Leu?Gln?Thr?Leu?Gln?Thr?Trp?Asn?Thr?Ser?Tyr?Pro

365 370 375 380

aag?cgc?agc?gcc?ggg?gag?gcc?ttc?atc?atc?tgc?ggc?acg?ctg?tac?gtc 1383

Lys?Arg?Ser?Ala?Gly?Glu?Ala?Phe?Ile?Ile?Cys?Gly?Thr?Leu?Tyr?Val

385 390 395

acc?aac?ggc?tac?tca?ggg?ggt?acc?aag?gtc?cac?tat?gca?tac?cag?acc 1431

Thr?Asn?Gly?Tyr?Ser?Gly?Gly?Thr?Lys?Val?His?Tyr?Ala?Tyr?Gln?Thr

400 405 410

aat?gcc?tcc?acc?tat?gaa?tac?atc?gac?atc?cca?ttc?cag?aac?aaa?tac 1479

Asn?Ala?Ser?Thr?Tyr?Glu?Tyr?Ile?Asp?Ile?Pro?Phe?Gln?Asn?Lys?Tyr

415 420 425

tcc?cac?atc?tcc?atg?ctg?gac?tac?aac?ccc?aag?gac?cgg?gcc?ctg?tat 1527

Ser?His?Ile?Ser?Met?Leu?Asp?Tyr?Asn?Pro?Lys?Asp?Arg?Ala?Leu?Tyr

430 435 440

gcc?tgg?aac?aac?ggc?cac?cag?atc?ctc?tac?aac?gtg?acc?ctc?ttc?cac 1575

Ala?Trp?Asn?Asn?Gly?His?Gln?Ile?Leu?Tyr?Asn?Val?Thr?Leu?Phe?His

445 450 455 460

gtc?atc?cgc?tcc?gac?gag?ttg?tag?ctccctcctc?ctggaagcca?agggcccacg 1629

Val?Ile?Arg?Ser?Asp?Glu?Leu

465

tcctcaccac?aaagggactc?ctgtgaaact?gctgccaaaa?agataccaat?aacactaaca 1689

ataccgatct?tgaaaaatca?tcagcagtgc?ggattctgac?atcgagggat?ggcattacct 1749

ccgtgtttct?ccctttcgag?ccggcgggcc?acagacgtcg?gaagaaactc?ccgtatttgc 1809

agctggaact?gcagcccacg?gcgccccggt?tttcctcccc?gccctgtccc?tctctggtca 1869

aacaacatac?taaagaggcg?aggcaatgac?tgttggccag?ttctcaccgg?ggaaaaaccc 1929

actgttagga?tggcatgaac?atttccttag?atcgtggtca?gctccgagga?atgtggcgtc 1989

caggctcttt?gagagccatg?ggctgcaccc?ggccgtaggc?tagtgtaact?cgcatcccat 2049

tgcagtgccg?tttcttgact?gtgttgctgt?ctcttagatt?aaccgtgctg?aggctccaca 2109

tagctcctgg?acctgtgtct?agtacatact?gaagcgatgg?tcagagtgtg?tagagtgaag 2169

ttgctgtgcc?cacattgttt?gaactcgcgt?accccgtaga?tacattgtgc?aacgttcttc 2229

tgttattccc?ttgaggtggt?aacttcgtat?gttcagttta?tgcgatgatt?gttgtaaatg 2289

caatgccgta?gtttggatta?ataagtggat?ggtttttgtt?tctaaaaaga?aaaaaaaaat 2349

cagtgttcac?ccttatagag?acatagtcaa?gttcatgttg?ataataatca?aaggaattac 2409

tctcttcttg?ttaaattagc?taaatcatgt?aaccgcagat?aggaagggct?cacctgggga 2469

aactctggtt?tccgatggga?caggaaagtc?atacgggcaa?cagtatgcgg?aaagtacgtt 2529

tttttaagta?aaaaacaaag?gcaaactttg?tactatccag?ttatctaagg?aacaataaaa 2589

acattaggag?aaaaaaaaaa?aaaaaaaaaa?aaaaaaaaaa?aaaaaaaaaa?aaaaaaaaaa 2649

aa 2651

<210>20

<211>467

<212>PRT

＜213〉people

<400>20

Met?Pro?Gly?Arg?Trp?Arg?Trp?Gln?Arg?Asp?Met?His?Pro?Ala?Arg?Lys

1 5 10 15

Leu?Leu?Ser?Leu?Leu?Phe?Leu?Ile?Leu?Met?Gly?Thr?Glu?Leu?Thr?Gln

20 25 30

Val?Leu?Pro?Thr?Asn?Pro?Glu?Glu?Ser?Trp?Gln?Val?Tyr?Ser?Ser?Ala

35 40 45

Gln?Asp?Ser?Glu?Gly?Arg?Cys?Ile?Cys?Thr?Val?Val?Ala?Pro?Gln?Gln

50 55 60

Thr?Met?Cys?Ser?Arg?Asp?Ala?Arg?Thr?Lys?Gln?Leu?Arg?Gln?Leu?Leu

65 70 75 80

Glu?Lys?Val?Gln?Asn?Met?Ser?Gln?Ser?Ile?Glu?Val?Leu?Asp?Arg?Arg

85 90 95

Thr?Gln?Arg?Asp?Leu?Gln?Tyr?Val?Glu?Lys?Met?Glu?Asn?Gln?Met?Lys

100 105 110

Gly?Leu?Glu?Ser?Lys?Phe?Lys?Gln?Val?Glu?Glu?Ser?His?Lys?Gln?His

115 120 125

Leu?Ala?Arg?Gln?Phe?Lys?Ala?Ile?Lys?Ala?Lys?Met?Asp?Glu?Leu?Arg

130 135 140

Pro?Leu?Ile?Pro?Val?Leu?Glu?Glu?Tyr?Lys?Ala?Asp?Ala?Lys?Leu?Val

145 150 155 160

Leu?Gln?Phe?Lys?Glu?Glu?Val?Gln?Asn?Leu?Thr?Ser?Val?Leu?Asn?Glu

165 170 175

Leu?Gln?Glu?Glu?Ile?Gly?Ala?Tyr?Asp?Tyr?Asp?Glu?Leu?Gln?Ser?Arg

180 185 190

Val?Ser?Asn?Leu?Glu?Glu?Arg?Leu?Arg?Ala?Cys?Met?Gln?Lys?Leu?Ala

195 200 205

Cys?Gly?Lys?Leu?Thr?Gly?Ile?Ser?Asp?Pro?Val?Thr?Val?Lys?Thr?Ser

210 215 220

Gly?Ser?Arg?Phe?Gly?Ser?Trp?Met?Thr?Asp?Pro?Leu?Ala?Pro?Glu?Gly

225 230 235 240

Asp?Asn?Arg?Val?Trp?Tyr?Met?Asp?Gly?Tyr?His?Asn?Asn?Arg?Phe?Val

245 250 255

Arg?Glu?Tyr?Lys?Ser?Met?Val?Asp?Phe?Met?Asn?Thr?Asp?Asn?Phe?Thr

260 265 270

Ser?His?Arg?Leu?Pro?His?Pro?Trp?Ser?Gly?Thr?Gly?Gln?Val?Val?Tyr

275 280 285

Asn?Gly?Ser?Ile?Tyr?Phe?Asn?Lys?Phe?Gln?Ser?His?Ile?Ile?Ile?Arg

290 295 300

Phe?Asp?Leu?Lys?Thr?Glu?Thr?Ile?Leu?Lys?Thr?Arg?Ser?Leu?Asp?Tyr

305 310 315 320

Ala?Gly?Tyr?Asn?Asn?Met?Tyr?His?Tyr?Ala?Trp?Gly?Gly?His?Ser?Asp

325 330 335

Ile?Asp?Leu?Met?Val?Asp?Glu?Ser?Gly?Leu?Trp?Ala?Val?Tyr?Ala?Thr

340 345 350

Asn?Gln?Asn?Ala?Gly?Asn?Ile?Val?Val?Ser?Arg?Leu?Asp?Pro?Val?Ser

355 360 365

Leu?Gln?Thr?Leu?Gln?Thr?Trp?Asn?Thr?Ser?Tyr?Pro?Lys?Arg?Ser?Ala

370 375 380

Gly?Glu?Ala?Phe?Ile?Ile?Cys?Gly?Thr?Leu?Tyr?Val?Thr?Asn?Gly?Tyr

385 390 395 400

Ser?Gly?Gly?Thr?Lys?Val?His?Tyr?Ala?Tyr?Gln?Thr?Asn?Ala?Ser?Thr

405 410 415

Tyr?Glu?Tyr?Ile?Asp?Ile?Pro?Phe?Gln?Asn?Lys?Tyr?Ser?His?Ile?Ser

420 425 430

Met?Leu?Asp?Tyr?Asn?Pro?Lys?Asp?Arg?Ala?Leu?Tyr?Ala?Trp?Asn?Asn

435 440 445

Gly?His?Gln?Ile?Leu?Tyr?Asn?Val?Thr?Leu?Phe?His?Val?Ile?Arg?Ser

450 455 460

Asp?Glu?Leu

465

<210>21

<211>2087

<212>DNA

＜213〉people

<220>

<221>CDS

<222>(106)..(1023)

<400>21

tttctccggg?aaccgtgccg?ggagagcgcg?cggtgctgga?gccgcaccgg?gtggccgaag 60

cagaagactt?tccggaagct?gctgggggat?gtctgactag?ctctc?atg?gag?ctc?cac 117

Met?Glu?Leu?His

1

tac?ctt?gct?aag?aag?agc?aac?cag?gca?gac?ctc?tgt?gat?gcc?agg?gac 165

Tyr?Leu?Ala?Lys?Lys?Ser?Asn?Gln?Ala?Asp?Leu?Cys?Asp?Ala?Arg?Asp

5 10 15 20

tgg?agt?tca?aga?ggg?ctg?cct?ggt?gac?cag?gca?gat?aca?gca?gcc?aca 213

Trp?Ser?Ser?Arg?Gly?Leu?Pro?Gly?Asp?Gln?Ala?Asp?Thr?Ala?Ala?Thr

25 30 35

aga?gct?gct?ctc?tgc?tgt?cag?aaa?cag?tgt?gca?tcc?acc?cca?aga?gca 261

Arg?Ala?Ala?Leu?Cys?Cys?Gln?Lys?Gln?Cys?Ala?Ser?Thr?Pro?Arg?Ala

40 45 50

acc?gag?atg?gaa?ggg?tct?aaa?ctt?agt?tct?tct?cca?gca?tcc?ccc?tcc 309

Thr?Glu?Met?Glu?Gly?Ser?Lys?Leu?Ser?Ser?Ser?Pro?Ala?Ser?Pro?Ser

55 60 65

tcc?tct?ctg?caa?aac?agt?act?ctt?cag?cca?gat?gcc?ttt?cca?cca?gga 357

Ser?Ser?Leu?Gln?Asn?Ser?Thr?Leu?Gln?Pro?Asp?Ala?Phe?Pro?Pro?Gly

70 75 80

ctt?ctc?cac?tca?ggg?aac?aac?caa?ata?aca?gcg?gaa?cgg?aaa?gtc?tgt 405

Leu?Leu?His?Ser?Gly?Asn?Asn?Gln?Ile?Thr?Ala?Glu?Arg?Lys?Val?Cys

85 90 95 100

aac?tgc?tgc?agc?cag?gaa?tta?gaa?act?tct?ttt?acc?tat?gtg?gac?aaa 453

Asn?Cys?Cys?Ser?Gln?Glu?Leu?Glu?Thr?Ser?Phe?Thr?Tyr?Val?Asp?Lys

105 110 115

aac?atc?aac?ttg?gag?cag?cgg?aac?cgg?agc?tcg?cca?tca?gca?aaa?ggg 501

Asn?Ile?Asn?Leu?Glu?Gln?Arg?Asn?Arg?Ser?Ser?Pro?Ser?Ala?Lys?Gly

120 125 130

cat?aat?cac?cct?ggg?gag?ctt?ggc?tgg?gaa?aat?cca?aat?gag?tgg?tcc 549

His?Asn?His?Pro?Gly?Glu?Leu?Gly?Trp?Glu?Asn?Pro?Asn?Glu?Trp?Ser

135 140 145

caa?gag?gct?gcc?ata?tct?ttg?ata?tct?gaa?gag?gag?gat?gat?aca?agt 597

Gln?Glu?Ala?Ala?Ile?Ser?Leu?Ile?Ser?Glu?Glu?Glu?Asp?Asp?Thr?Ser

150 155 160

tca?gaa?gcc?acg?tct?tca?ggg?aag?tct?ata?gac?tat?ggt?ttc?atc?agc 645

Ser?Glu?Ala?Thr?Ser?Ser?Gly?Lys?Ser?Ile?Asp?Tyr?Gly?Phe?Ile?Ser

165 170 175 180

gcc?atc?ttg?ttc?ttg?gtc?act?ggg?atc?ctg?ctc?gtg?atc?atc?tct?tac 693

Ala?Ile?Leu?Phe?Leu?Val?Thr?Gly?Ile?Leu?Leu?Val?Ile?Ile?Ser?Tyr

185 190 195

atc?gtc?cca?cgg?gaa?gtg?act?gtg?gac?ccc?aac?act?gtg?gca?gcc?cgg 741

Ile?Val?Pro?Arg?Glu?Val?Thr?Val?Asp?Pro?Asn?Thr?Val?Ala?Ala?Arg

200 205 210

gag?atg?gag?cgc?ctg?gag?aag?gag?agt?gcg?agg?ctg?ggg?gct?cac?ctg 789

Glu?Met?Glu?Arg?Leu?Glu?Lys?Glu?Ser?Ala?Arg?Leu?Gly?Ala?His?Leu

215 220 225

gac?cgc?tgt?gtg?att?gcg?ggg?ctc?tgc?ctc?ctc?acg?ctg?ggg?ggc?gtc 837

Asp?Arg?Cys?Val?Ile?Ala?Gly?Leu?Cys?Leu?Leu?Thr?Leu?Gly?Gly?Val

230 235 240

atc?ctg?tcc?tgc?ttg?tta?atg?atg?tcc?atg?tgg?aag?ggg?gag?ctc?tat 885

Ile?Leu?Ser?Cys?Leu?Leu?Met?Met?Ser?Met?Trp?Lys?Gly?Glu?Leu?Tyr

245 250 255 260

cgt?cga?aac?aga?ttt?gcc?tct?tcc?aaa?gag?tct?gca?aaa?ctc?tat?ggt 933

Arg?Arg?Asn?Arg?Phe?Ala?Ser?Ser?Lys?Glu?Ser?Ala?Lys?Leu?Tyr?Gly

265 270 275

tct?ttc?aac?ttc?agg?atg?aaa?acc?agc?acg?aat?gaa?aac?act?ctg?gaa 981

Ser?Phe?Asn?Phe?Arg?Met?Lys?Thr?Ser?Thr?Asn?Glu?Asn?Thr?Leu?Glu

280 285 290

ctg?tcc?ttg?gta?gag?gaa?gat?gcg?ctt?gct?gta?cag?agt?taa 1023

Leu?Ser?Leu?Val?Glu?Glu?Asp?Ala?Leu?Ala?Val?Gln?Ser

295 300 305

ttctggttgt?gaatatcttg?agagtctgcc?ttggcatttt?ataatatgaa?aaaagttaat 1083

ttataaaaat?tcacagtgca?atttatttgc?ctggcaagaa?aagtttattt?cacaaaccaa 1143

cagccagtaa?gtgtttttgt?tctctatgtg?tcttctattt?agaagaaaag?ccatgtaaga 1203

tgtataagaa?accacaacca?gccacaccta?tccttctgaa?gagctgaagg?ctaattaatc 1263

tgtaatggcc?aagaacttct?acttcgatag?aaaaatattt?ctaatgaccc?agtctacaaa 1323

ttatttcttt?tacacaaata?tatgatgtta?ttctttggac?actaggtggt?cctacacaca 1383

gtaggatcaa?ttgctaatct?actttgtgaa?aaagaactaa?gcactaatca?ataataaggc 1443

ttacatctaa?ttctcaaagg?tgcttatcca?ttttcttgct?aaattatcct?tcttgtaatt 1503

tggctaaaca?ctaaaacatg?gaatttttag?tttgaatatt?ttgaagtttg?aggatgttgg 1563

gctttcctta?ttgtaaaaaa?tgttatgttt?gaaattattc?ctgttttcaa?aaatggtaat 1623

taagtcatta?ggataaactt?tctaataaaa?aaaaattatg?taaatcattg?taagaccaat 1683

gtgaatttaa?accacagtgt?tggtgccaga?tattagccag?ggtcacaggc?tcttttttaa 1743

aatgaagtca?aattagttac?tgatataagt?tttatatttt?gtgattttgc?catcatactg 1803

gttatctgtg?ctcttgaagt?ctgctggttc?tcacaccagc?agcaagaggc?aaggcatgag 1863

acattttaaa?tatatttcca?gacatctaaa?catctagaca?ggcctgagtc?atgattggct 1923

ctggtgtgct?tccaggtaac?atcttgttat?tgctgtttgg?ggaaaaccac?caatagaatt 1983

gcagctagac?ttttggggac?tgtgcctcgc?aggctgtatc?acttgctgta?ggccaagcaa 2043

cgagcacttc?cctcatggta?gcaacagtcc?ttaagcccaa?agcc 2087

<210>22

<211>305

<212>PRT

＜213〉people

<400>22

Met?Glu?Leu?His?Tyr?Leu?Ala?Lys?Lys?Ser?Asn?Gln?Ala?Asp?Leu?Cys

1 5 10 15

Asp?Ala?Arg?Asp?Trp?Ser?Ser?Arg?Gly?Leu?Pro?Gly?Asp?Gln?Ala?Asp

20 25 30

Thr?Ala?Ala?Thr?Arg?Ala?Ala?Leu?Cys?Cys?Gln?Lys?Gln?Cys?Ala?Ser

35 40 45

Thr?Pro?Arg?Ala?Thr?Glu?Met?Glu?Gly?Ser?Lys?Leu?Ser?Ser?Ser?Pro

50 55 60

Ala?Ser?Pro?Ser?Ser?Ser?Leu?Gln?Asn?Ser?Thr?Leu?Gln?Pro?Asp?Ala

65 70 75 80

Phe?Pro?Pro?Gly?Leu?Leu?His?Ser?Gly?Asn?Asn?Gln?Ile?Thr?Ala?Glu

85 90 95

Arg?Lys?Val?Cys?Asn?Cys?Cys?Ser?Gln?Glu?Leu?Glu?Thr?Ser?Phe?Thr

100 105 110

Tyr?Val?Asp?Lys?Asn?Ile?Asn?Leu?Glu?Gln?Arg?Asn?Arg?Ser?Ser?Pro

115 120 125

Ser?Ala?Lys?Gly?His?Asn?His?Pro?Gly?Glu?Leu?Gly?Trp?Glu?Asn?Pro

130 135 140

Asn?Glu?Trp?Ser?Gln?Glu?Ala?Ala?Ile?Ser?Leu?Ile?Ser?Glu?Glu?Glu

145 150 155 160

Asp?Asp?Thr?Ser?Ser?Glu?Ala?Thr?Ser?Ser?Gly?Lys?Ser?Ile?Asp?Tyr

165 170 175

Gly?Phe?Ile?Ser?Ala?Ile?Leu?Phe?Leu?Val?Thr?Gly?Ile?Leu?Leu?Val

180 185 190

Ile?Ile?Ser?Tyr?Ile?Val?Pro?Arg?Glu?Val?Thr?Val?Asp?Pro?Asn?Thr

195 200 205

Val?Ala?Ala?Arg?Glu?Met?Glu?Arg?Leu?Glu?Lys?Glu?Ser?Ala?Arg?Leu

210 215 220

Gly?Ala?His?Leu?Asp?Arg?Cys?Val?Ile?Ala?Gly?Leu?Cys?Leu?Leu?Thr

225 230 235 240

Leu?Gly?Gly?Val?Ile?Leu?Ser?Cys?Leu?Leu?Met?Met?Ser?Met?Trp?Lys

245 250 255

Gly?Glu?Leu?Tyr?Arg?Arg?Asn?Arg?Phe?Ala?Ser?Ser?Lys?Glu?Ser?Ala

260 265 270

Lys?Leu?Tyr?Gly?Ser?Phe?Asn?Phe?Arg?Met?Lys?Thr?Ser?Thr?Asn?Glu

275 280 285

Asn?Thr?Leu?Glu?Leu?Ser?Leu?Val?Glu?Glu?Asp?Ala?Leu?Ala?Val?Gln

290 295 300

Ser

305

<210>23

<211>1882

<212>DNA

＜213〉people

<220>

<221>CDS

<222>(534)..(1505)

<400>23

ggcacgaggg?ttagattagt?ctgaagccgc?caccagcccc?aggcccccgt?gcagaagaaa 60

agcgggaggg?aacggcggag?gccgccgctg?ccctgcaccg?ccctcctgga?ggccacttgg 120

agagtccggc?cccgaggagg?ccatggccac?aagtgcccac?agctggcccc?aggttgccag 180

cgtcgctaca?gcccagacca?aggcagaata?atctccggat?gagctggtgg?caccgctgag 240

cctttggtct?caccagggct?tcctgttgct?ggcaggcggg?gtggagcgga?gctgctggga 300

ggctgctgga?taggagaggg?gtcacggctg?cggaagagga?ggttcttcgg?gacacccgtg 360

gatggacacg?gcaaggaaac?accaggccaa?ccacagctgg?ggataaaata?gcacaaccac 420

accctgccgt?ccagcgcctc?ccagcctgtg?ccccttccta?gtaccaccag?caaccatcaa 480

tcccgtctcc?tcctgcctcc?tctcctgcaa?tccaccccgc?cacgactatc?gcc?atg 536

Met

1

gca?gcc?ctg?atc?gca?gag?aac?ttc?cgc?ttc?ctg?tca?ctt?ttc?ttc?aag 584

Ala?Ala?Leu?Ile?Ala?Glu?Asn?Phe?Arg?Phe?Leu?Ser?Leu?Phe?Phe?Lys

5 10 15

agc?aag?gat?gtg?atg?att?ttc?aac?ggc?ctg?gtg?gca?ctg?ggc?acg?gtg 632

Ser?Lys?Asp?Val?Met?Ile?Phe?Asn?Gly?Leu?Val?Ala?Leu?Gly?Thr?Val

20 25 30

ggc?agc?cag?gag?ctg?ttc?tct?gtg?gtg?gcc?ttc?cac?tgc?ccc?tgc?tcg 680

Gly?Ser?Gln?Glu?Leu?Phe?Ser?Val?Val?Ala?Phe?His?Cys?Pro?Cys?Ser

35 40 45

ccg?gcc?cgg?aac?tac?ctg?tac?ggg?ctg?gcg?gcc?atc?ggc?gtg?ccc?gcc 728

Pro?Ala?Arg?Asn?Tyr?Leu?Tyr?Gly?Leu?Ala?Ala?Ile?Gly?Val?Pro?Ala

50 55 60 65

ctg?gtg?ctc?ttc?atc?att?ggc?atc?atc?ctc?aac?aac?cac?acc?tgg?aac 776

Leu?Val?Leu?Phe?Ile?Ile?Gly?Ile?Ile?Leu?Asn?Asn?His?Thr?Trp?Asn

70 75 80

ctc?gtg?gcc?gag?tgc?cag?cac?cgg?agg?acc?aag?aac?tgc?tcc?gcc?gcc 824

Leu?Val?Ala?Glu?Cys?Gln?His?Arg?Arg?Thr?Lys?Asn?Cys?Ser?Ala?Ala

85 90 95

ccc?acc?ttc?ctc?ctt?cta?agc?tcc?atc?ctg?gga?cgt?gcg?gct?gtg?gcc 872

Pro?Thr?Phe?Leu?Leu?Leu?Ser?Ser?Ile?Leu?Gly?Arg?Ala?Ala?Val?Ala

100 105 110

cct?gtc?acc?tgg?tct?gtc?atc?tcc?ctg?ctg?cgt?ggt?gag?gct?tat?gtc 920

Pro?Val?Thr?Trp?Ser?Val?Ile?Ser?Leu?Leu?Arg?Gly?Glu?Ala?Tyr?Val

115 120 125

tgt?gct?ctc?agt?gag?ttc?gtg?gac?cct?tcc?tca?ctc?acg?gcc?agg?gaa 968

Cys?Ala?Leu?Ser?Glu?Phe?Val?Asp?Pro?Ser?Ser?Leu?Thr?Ala?Arg?Glu

130 135 140 145

gag?cacttc?cca?tca?gcc?cac?gcc?act?gaa?atc?ctg?gcc?agg?ttc?ccc 1016

Glu?His?Phe?Pro?Ser?Ala?His?Ala?Thr?Glu?Ile?Leu?Ala?Arg?Phe?Pro

150 155 160

tgc?aag?gag?aac?cct?gac?aac?ctg?tca?gac?ttc?cgg?gag?gag?gtc?agc 1064

Cys?Lys?Glu?Asn?Pro?Asp?Asn?Leu?Ser?Asp?Phe?Arg?Glu?Glu?Val?Ser

165 170 175

cgc?agg?ctc?agg?tat?gag?tcc?cag?ctc?ttt?gga?tgg?ctg?ctc?atc?ggc 1112

Arg?Arg?Leu?Arg?Tyr?Glu?Ser?Gln?Leu?Phe?Gly?Trp?Leu?Leu?Ile?Gly

180 185 190

gtg?gtg?gcc?atc?ctg?gtg?ttc?ctg?acc?aag?tgc?ctc?aag?cat?tac?tgc 1160

Val?Val?Ala?Ile?Leu?Val?Phe?Leu?Thr?Lys?Cys?Leu?Lys?His?Tyr?Cys

195 200 205

tca?cca?ctc?agc?tac?cgc?cag?gag?gcc?tac?tgg?gcg?cag?tac?cgc?gcc 1208

Ser?Pro?Leu?Ser?Tyr?Arg?Gln?Glu?Ala?Tyr?Trp?Ala?Gln?Tyr?Arg?Ala

210 215 220 225

aat?gag?gac?cag?ctg?ttc?cag?cgc?acg?gcc?gag?gtg?cac?tct?cgg?gtg 1256

Asn?Glu?Asp?Gln?Leu?Phe?Gln?Arg?Thr?Ala?Glu?Val?His?Ser?Arg?Val

230 235 240

ctc?gct?gcc?aac?aat?gtg?cgc?cgc?ttc?ttt?ggc?ttt?gtg?gcg?ctc?aac 1304

Leu?Ala?Ala?Asn?Asn?Val?Arg?Arg?Phe?Phe?Gly?Phe?Val?Ala?Leu?Asn

245 250 255

aag?gat?gat?gag?gaa?ctg?att?gcc?aac?ttc?cca?gtg?gaa?ggc?acg?cag 1352

Lys?Asp?Asp?Glu?Glu?Leu?Ile?Ala?Asn?Phe?Pro?Val?Glu?Gly?Thr?Gln

260 265 270

cca?cgg?cca?cag?tgg?aat?gcc?atc?acc?ggc?gtc?tac?ttg?tac?cgt?gag 1400

Pro?Arg?Pro?Gln?Trp?Asn?Ala?Ile?Thr?Gly?Val?Tyr?Leu?Tyr?Arg?Glu

275 280 285

aac?cag?ggc?ctc?cca?ctc?tac?agc?cgc?ctg?cac?aag?tgg?gcc?cag?ggt 1448

Asn?Gln?Gly?Leu?Pro?Leu?Tyr?Ser?Arg?Leu?His?Lys?Trp?Ala?Gln?Gly

290 295 300 305

ctg?gca?ggc?aac?ggc?gcg?gcc?cct?gac?aac?gtg?gag?atg?gcc?ctg?ctc 1496

Leu?Ala?Gly?Asn?Gly?Ala?Ala?Pro?Asp?Asn?Val?Glu?Met?Ala?Leu?Leu

310 315 320

ccc?tcc?taa?ggaggtgctt?cccatgctct?ttgtaaatgg?cactacttgg 1545

Pro?Ser

tcccaaactg?aaccccactg?cttgctcaca?tccatatcag?aaggggattt?ttaaaaaact 1605

gttatcttct?tggccagggg?aaaggaccac?aaggcaatct?ggggtgtgga?cagacccagt 1665

agacaatgga?agccccagcc?agcagggcca?ggtgacagtg?aagctcacca?gtgggctcct 1725

ttatggtact?ctatgcagtt?aacatgtatc?tagctgcata?gggacaccca?gcgcagcagt 1785

gcaccactgg?gaagtggcct?ccagtgcagc?ctctggcctt?attttatata?tttaaatttt 1845

tgataaagtt?tttcttacta?aaaggaaaaa?aaaaaaa 1882

<210>24

<211>323

<212>PRT

＜213〉people

<400>24

Met?Ala?Ala?Leu?Ile?Ala?Glu?Asn?Phe?Arg?Phe?Leu?Ser?Leu?Phe?Phe

1 5 10 15

Lys?Ser?Lys?Asp?Val?Met?Ile?Phe?Asn?Gly?Leu?Val?Ala?Leu?Gly?Thr

20 25 30

Val?Gly?Ser?Gln?Glu?Leu?Phe?Ser?Val?Val?Ala?Phe?His?Cys?Pro?Cys

35 40 45

Ser?Pro?Ala?Arg?Asn?Tyr?Leu?Tyr?Gly?Leu?Ala?Ala?Ile?Gly?Val?Pro

50 55 60

Ala?Leu?Val?Leu?Phe?Ile?Ile?Gly?Ile?Ile?Leu?Asn?Asn?His?Thr?Trp

65 70 75 80

Asn?Leu?Val?Ala?Glu?Cys?Gln?His?Arg?Arg?Thr?Lys?Asn?Cys?Ser?Ala

85 90 95

Ala?Pro?Thr?Phe?Leu?Leu?Leu?Ser?Ser?Ile?Leu?Gly?Arg?Ala?Ala?Val

100 105 110

Ala?Pro?Val?Thr?Trp?Ser?Val?Ile?Ser?Leu?Leu?Arg?Gly?Glu?Ala?Tyr

115 120 125

Val?Cys?Ala?Leu?Ser?Glu?Phe?Val?Asp?Pro?Ser?Ser?Leu?Thr?Ala?Arg

130 135 140

Glu?Glu?His?Phe?Pro?Ser?Ala?His?Ala?Thr?Glu?Ile?Leu?Ala?Arg?Phe

145 150 155 160

Pro?Cys?Lys?Glu?Asn?Pro?Asp?Asn?Leu?Ser?Asp?Phe?Arg?Glu?Glu?Val

165 170 175

Ser?Arg?Arg?Leu?Arg?Tyr?Glu?Ser?Gln?Leu?Phe?Gly?Trp?Leu?Leu?Ile

180 185 190

Gly?Val?Val?Ala?Ile?Leu?Val?Phe?Leu?Thr?Lys?Cys?Leu?Lys?His?Tyr

195 200 205

Cys?Ser?Pro?Leu?Ser?Tyr?Arg?Gln?Glu?Ala?Tyr?Trp?Ala?Gln?Tyr?Arg

210 215 220

Ala?Asn?Glu?Asp?Gln?Leu?Phe?Gln?Arg?Thr?Ala?Glu?Val?His?Ser?Arg

225 230 235 240

Val?Leu?Ala?Ala?Asn?Asn?Val?Arg?Arg?Phe?Phe?Gly?Phe?Val?Ala?Leu

245 250 255

Asn?Lys?Asp?Asp?Glu?Glu?Leu?Ile?Ala?Asn?Phe?Pro?Val?Glu?Gly?Thr

260 265 270

Gln?Pro?Arg?Pro?Gln?Trp?Asn?Ala?Ile?Thr?Gly?Val?Tyr?Leu?Tyr?Arg

275 280 285

Glu?Asn?Gln?Gly?Leu?Pro?Leu?Tyr?Ser?Arg?Leu?His?Lys?Trp?Ala?Gln

290 295 300

Gly?Leu?Ala?Gly?Asn?Gly?Ala?Ala?Pro?Asp?Asn?Val?Glu?Met?Ala?Leu

305 310 315 320

Leu?Pro?Ser

<210>25

<211>673

<212>DNA

＜213〉people

<220>

<221>CDS

<222>(135)..(467)

<400>25

cttccgagca?agatggcgcc?gcgggcattt?cttccactgc?ccgtctgagg?gaacgctaag 60

tagtgtgtcc?ggcgccgtgt?tccagctccg?cgttgttccg?cgagaaagcg?agaggccgag 120

cctgggctgg?tgcg?atg?gcc?gcg?gtg?gtg?gcc?aag?cgg?gaa?ggg?ccg?ccg 170

Met?Ala?Ala?Val?Val?Ala?Lys?Arg?Glu?Gly?Pro?Pro

1 5 10

ttc?atc?agc?gag?gcg?gcc?gtg?cgg?ggc?aac?gcc?gcc?gtc?ctg?gat?tat 218

Phe?Ile?Ser?Glu?Ala?Ala?Val?Arg?Gly?Asn?Ala?Ala?Val?Leu?Asp?Tyr

15 20 25

tgc?cgg?acc?tcg?gtg?tca?gcg?ctg?tcg?ggg?gcc?acg?gcc?ggc?atc?ctc 266

Cys?Arg?Thr?Ser?Val?Ser?Ala?Leu?Ser?Gly?Ala?Thr?Ala?Gly?Ile?Leu

30 35 40

ggc?ctc?acc?ggc?ctc?tac?ggc?ttc?atc?ttc?tac?ctg?ctc?gcc?tcc?gtc 314

Gly?Leu?Thr?Gly?Leu?Tyr?Gly?Phe?Ile?Phe?Tyr?Leu?Leu?Ala?Ser?Val

45 50 55 60

ctg?ctc?tcc?ctg?ctc?ctc?att?ctc?aag?gcg?gga?agg?agg?tgg?aac?aaa 362

Leu?Leu?Ser?Leu?Leu?Leu?Ile?Leu?Lys?Ala?Gly?Arg?Arg?Trp?Asn?Lys

65 70 75

tat?ttc?aaa?tca?cgg?aga?cct?ctc?ttt?aca?gga?ggc?ctc?atc?ggg?ggc 410

Tyr?Phe?Lys?Ser?Arg?Arg?Pro?Leu?Phe?Thr?Gly?Gly?Leu?Ile?Gly?Gly

80 85 90

ctc?ttc?acc?tac?gtc?ctg?ttc?tgg?acg?ttc?ctc?tac?ggc?atg?gtg?cac 458

Leu?Phe?Thr?Tyr?Val?Leu?Phe?Trp?Thr?Phe?Leu?Tyr?Gly?Met?Val?His

95 100 105

gtc?tac?tga?aatgggggcc?cgggggactt?ttttaaaaaa?ccagatcggg 507

Val?Tyr

110

aggactgtgg?ccagcaatta?acaccatgta?gacttcctta?gttcttaagt?ggttgaattc 567

gctgcttgtt?ctgtaacgtt?ataaataatt?tatatctgaa?gacggagagc?ctgtaatatt 627

cttcagatta?aatgaagcgt?gagacaaaaa?aaaaaaaaaa?aaaaaa 673

<210>26

<211>110

<212>PRT

＜213〉people

<400>26

Met?Ala?Ala?Val?Val?Ala?Lys?Arg?Glu?Gly?Pro?Pro?Phe?Ile?Ser?Glu

1 5 10 15

Ala?Ala?Val?Arg?Gly?Asn?Ala?Ala?Val?Leu?Asp?Tyr?Cys?Arg?Thr?Ser

20 25 30

Val?Ser?Ala?Leu?Ser?Gly?Ala?Thr?Ala?Gly?Ile?Leu?Gly?Leu?Thr?Gly

35 40 45

Leu?Tyr?Gly?Phe?Ile?Phe?Tyr?Leu?Leu?Ala?Ser?Val?Leu?Leu?Ser?Leu

50 55 60

Leu?Leu?Ile?Leu?Lys?Ala?Gly?Arg?Arg?Trp?Asn?Lys?Tyr?Phe?Lys?Ser

65 70 75 80

Arg?Arg?Pro?Leu?Phe?Thr?Gly?Gly?Leu?Ile?Gly?Gly?Leu?Phe?Thr?Tyr

85 90 95

Val?Leu?Phe?Trp?Thr?Phe?Leu?Tyr?Gly?Met?Val?His?Val?Tyr

100 105 110

Claims (16)

1. isolating polynucleotide is characterized in that, described polynucleotide contain the nucleotide sequence that coding has the polypeptide of cell death inducing function, and this nucleotide sequence is selected from:

(a) polynucleotide of polypeptide that contain the aminoacid sequence of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ IDNO:22, SEQ ID NO:24, SEQ ID NO:26 with coding have the polynucleotide of at least 70% similarity;

(b) coding contains the polynucleotide of polypeptide that aminoacid sequence with SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ IDNO:22, SEQ ID NO:24, SEQ ID NO:26 has the aminoacid sequence of at least 70% similarity;

(c) with (a) or polynucleotide complementary polynucleotide (b).

2. polynucleotide as claimed in claim 1, it is characterized in that the polypeptide of described polynucleotide encoding has the aminoacid sequence of the group of being selected from down: SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ IDNO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26.

3. polynucleotide as claimed in claim 1 is characterized in that, the sequence of described polynucleotide is shown at least 85% similarity with the nucleotides sequence that is selected from down group:

(a) coding region sequence or the full length sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ IDNO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25;

(b) at least one sequence of the sequence of in genetic code degeneracy scope, mentioning in corresponding to (a);

(c) with (a) or at least one sequence of the sequence complementary sequence hybridization of mentioning (b).

4. polynucleotide as claimed in claim 3, it is characterized in that the sequence of described polynucleotide is selected from coding region sequence or the full length sequence of SEQ ID NO:1, SEQ IDNO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ IDNO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25.

5. the polypeptide of the described polynucleotide encoding of claim 1, it is characterized in that described polypeptide comprises the polypeptide that is selected from down the aminoacid sequence in the group: SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26; Or the polypeptide that has at least 90% similarity with above arbitrary aminoacid sequence; Or its conservative property variation polypeptide or its active fragments or its reactive derivative.

6. the described polypeptide of claim 5, it is characterized in that described polypeptide has the aminoacid sequence of the group of being selected from down: SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26.

7. a carrier is characterized in that, described carrier contains the described polynucleotide of claim 1.

8. a genetically engineered host cell is characterized in that, described host cell is selected from:

(a) host cell that transforms or transduce with the described carrier of claim 7;

(b) host cell that transforms or transduce with the described polynucleotide of claim 1.

9. an antibody is characterized in that, described antibody be can with the described polypeptid specificity bonded of claim 5 antibody.

10. a nucleic acid fragment is characterized in that, described nucleic acid fragment contains 8-100 successive Nucleotide in the described arbitrary polynucleotide of claim 1.

11. the application of the described polynucleotide of claim 1 in cell death inducing, heterogenous expression can cell death inducing in host cell to it is characterized in that described polynucleotide.

12. the described application of claim 11 is characterized in that described host cell is selected from the described host cell of claim 8.

13. described polynucleotide of claim 1 or the described polypeptide of claim 5 prevent and/or treat purposes in the medicine with human body cell apoptosis diseases associated in preparation.

14. purposes according to claim 13, wherein said disease is selected from as autoimmune disease and tumours such as acquired immune deficiency syndrome (AIDS), neural degeneration.

15. contain the pharmaceutical composition of described polypeptide of claim 5 and pharmaceutically acceptable carrier.

16. the external detection method of autoimmune disease or tumour is characterized in that, utilizes described antibody of claim 9 or claim 10 described nucleic acid fragments to detect the existence or the level of the polypeptide in host's sample.

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